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Purpose It is of great significance to accurately identify the KRAS gene mutation status for patients in tumor prognosis and personalized treatment. Although the computer-aided diagnosis system based on deep learning has gotten all-round development, its performance still cannot meet the current clinical application requirements due to the inherent limitations of small-scale medical image dataset and inaccurate lesion feature extraction. Therefore, our aim is to propose a deep learning model based on T2 MRI of colorectal cancer patients to identify whether KRAS gene is mutated. Methods In this research, a multi-task attentive model is proposed to identify KRAS gene mutations in patients, which is mainly composed of a segmentation sub-network and an identification sub-network. Specifically, at first, the features extracted by the encoder of segmentation model are used as guidance information to guide the two attention modules in the identification network for precise activation of the lesion area. Then the original image of the lesion and the segmentation result are concatenated for feature extraction. Finally, features extracted from the second step are combined with features activated by the attention modules to identify the gene mutation status. In this process, we introduce the inter-layer loss function to encourage the similarity of the two sub-network parameters and ensure that the key features are fully extracted to alleviate the overfitting problem caused by small dataset to some extent. Results The proposed identification model is benchmarked primarily using 15-fold cross validation. 382 images from 36 clinical cases were used to test the model. For the identification of KRAS mutation status, the average accuracy is 89.95±1.23%, the average sensitivity is 89.29±1.79%, the average specificity is 90.53±2.45%, and the average AUC is 95.73±5.02%. For segmentation of lesions, the average dice is 88.11 ± 0.86% CONCLUSIONS: We developed a novel deep learning-based model to identify the KRAS status in colorectal cancer. We demonstrated the excellent properties of the proposed identification through comparison with ground truth gene mutation status of 36 clinical cases. And all these results show that the novel method has great potential for clinical application. This article is protected by copyright. All rights reserved.

Background Pancreatic cancer (PC) is one of the deadliest malignancies with an alarming mortality rate. Despite significant advancement in diagnostics and therapeutics, early diagnosis remains elusive causing poor prognosis, marred by mutations and epigenetic modifications in key genes which contribute to disease progression. Aim To evaluate the various biological tumor markers collectively for early diagnosis which could act as prognostic biomarkers and helps in future therapeutics of PC in Kashmir valley. Methods A total of 50 confirmed PC cases were included in the study to evaluate the levels of carbohydrate antigen 19-9 (CA 19-9), tissue polypeptide specific antigen (TPS), carcinoembryonic antigen (CEA), vascular endothelial growth factor-A (VEGF-A), and epidermal growth factor receptor (EGFR). Mutational analysis was performed to evaluate the mutations in Kirsten rat sarcoma (KRAS), Breast cancer type 2 (BRCA-2), and deleted in pancreatic cancer-4 (DPC-4) genes. However, epigenetic modifications (methylation of CpG islands) were performed in the promoter regions of cyclin-dependent kinase inhibitor 2A (p16; CDKN2A), MutL homolog 1 (hMLH1), and Ras association domain-containing protein 1(RASSF1A) genes. Results We found significantly elevated levels of biological markers CA 19-9 (P ≤ 0.05), TPS (P ≤ 0.05), CEA (P ≤ 0.001), and VEGF (P ≤ 0.001). Molecular genetic analysis revealed that KRAS gene mutation is predominant in codon 12 (16 subjects, P ≤ 0.05), and 13 (12 subjects, P ≤ 0.05). However, we did not find a mutation in DPC-4 (1203G > T) and BRCA-2 (617delT) genes. Furthermore, epigenetic modification revealed that CpG methylation in 21 (P ≤ 0.05) and 4 subjects in the promoter regions of the p16 and hMLH1 gene, respectively. Conclusion In conclusion, CA 19-9, TPS, CEA, and VEGF levels were significantly elevated and collectively have potential as diagnostic and prognostic markers in PC. Global data of mutation in the KRAS gene commonly in codon 12 and rare in codon 13 could augment the predisposition towards PC. Additionally, methylation of the p16 gene could also modulate transcription of genes thereby increasing the predisposition and susceptibility towards PC.

Identifying Kirsten rat sarcoma (KRAS) mutation status is important for metastatic colorectal cancer treatment. Validating KRAS mutation profile on neuroimaging without histopathologic analysis would be beneficial. We aimed to differentiate Kirsten rat sarcoma (KRAS) gene mutation profile according to diffusion tensor imaging (DTI) metrics obtained from colorectal brain metastases (CRBM). Total of 8 patients with 17 CRBM included with known KRAS gene mutation status. All patients were categorized into two groups according to the KRAS mutation status as positive (+) (n: 5, 11 lesions) and negative (-) (wild type) (n: 3, 6 lesions). DTI values of the BMs were evaluated by consensus of two radiologists. Apparent diffusion coefficient (ADC), normalized apparent diffusion coefficient (nADC) and fractional anisotropy (FA) values of the two groups were compared. For comparison the Mann-Whitney U test was used. KRAS mutation (+) group showed significantly lower ADC and nADC values on CRBM rather than the wild-type. Comparison of the FA values of the two groups did not reveal any statistical significance. In conclusion, decreased ADC and nADC values of CRBM are correlated with presence of KRAS mutation. Lower ADC and nADC values obtained from CRBM can be utilized to insight KRAS mutation profile. [Med-Science 2021; 10(3.000): 886-9]

Background Colorectal cancer (CRC) is among the most widespread malignancies in the world. MicroRNA (miRNA) has been identified as an important modulator of the biological processes of the cells. This group of noncoding RNAs also has a pivotal function in the growth and development of human cancers, including CRC. Among these miRNAs, miR-196, miR-132, miR-146a, and miR-134 have fundamental impacts on the regulation of cancers. The current study aimed to investigate the involvement of these miRNAs in CRC patients. Methods In this study, 50 pairs of tumor and tumor margin samples of CRC patients were investigated to assess the expression levels of miR-196, miR-132, miR-146a, and miR-134 in this cancer. For this purpose, firstly, quantitative real-time PCR (qRT-PCR) was applied. Also, KRAS mutation and clinicopathological characteristics of the CRC patients were analyzed in the study groups. Results The findings demonstrated the overexpression of miR-196 (P-value = 0.0045) and miR-146a (P-value = 0.0033) in tumor tissues compared to controls. Conversely, the expression levels of miR-132 (P-value = 0.00032) and miR-134 (P-value < 0.0001) were downregulated in tumor tissues. Also, miR-146a was the only miRNA with significant expression change in the case of the KRAS gene mutation. Interestingly, the expression ratio of these miRNAs was significantly associated with some of the clinicopathological features of the patients, such as lymph node and distant metastases. Conclusion Our data demonstrated that these miRNAs appear to be promising novel biomarkers for early diagnosis of CRC and may pave the way for the future establishment of novel therapeutic options for CRC.

Background To analyze and evaluate EGFR, KRAS, and PIK3CA gene mutation rates and clinical distribution in patients with different types of lung cancer Method A total of 221 lung cancer patients treated in our hospital between January 2016 and June 2019 were enrolled. Tissue and whole blood samples were collected and analyzed to determine the mutation status of EGFR, KRAS, and PIK3CA genes. The gene exon mutation rates were determined. Relevant clinical data, such as age, gender, tumor sample type, treatment method, pathologic type, and lung cancer stage were recorded and statistically analyzed. Results The EGFR gene mutation rates in exons E18-E21 were 2.3%, 17.6%, 3.6%, and 20.4%, respectively. E18, E19, and E20 mutations were commonly detected in adenosquamous carcinoma, and E21 mutations were commonly detected in adenocarcinoma. Mutations in exons E18-E21 were frequently detected in patients with lung cancer stages IA, IB, IIA, or IIB, respectively. The KRAS gene mutation rate in lung cancer patients in exon E2 was higher in whole blood and tissue samples than other exon mutations, while the KRAS gene mutation rate in exons E2 and E3 was significantly higher in patients with lung cancer stages IIB and IA, respectively. PIK3CA gene mutations in exons E9 and E20 occurred in patients < 60 years of age. Exon E9-positive mutations were more common in men or patients with squamous cell carcinoma, while exon E20-positive mutations were more common in females. Conclusion The EGFR, KRAS, and PIK3CA gene exon mutation rates differ and were shown to be correlated with different clinical indicators, which have significance in clinical treatment.

Mutational activation of the KRAS gene occurs in almost all pancreatic ductal adenocarcinoma (PDAC) and is the earliest molecular event in their carcinogenesis. Evidence has accumulated of the metabolic reprogramming in PDAC, such as amino acid homeostasis and autophagic flux. However, the biological effects of KRAS mutation on metabolic reprogramming at the earlier stages of PDAC carcinogenesis are unclear. Here we report dynamic metabolic reprogramming in immortalized human non-cancerous pancreatic ductal epithelial cells, in which a KRAS mutation was induced by gene-editing, which may mimic early pancreatic carcinogenesis. Similar to the cases of PDAC, KRAS gene mutation increased the dependency on glucose and glutamine for maintaining the intracellular redox balance. In addition, the intracellular levels of amino acids were significantly decreased because of active protein synthesis, and the cells required greater autophagic flux to maintain their viability. The lysosomal inhibitor chloroquine significantly inhibited cell proliferation. Therefore, metabolic reprogramming is an early event in carcinogenesis initiated by KRAS gene mutation, suggesting a rationale for the development of nutritional interventions that suppress or delay the development of PDAC.

Colorectal cancer (CRC) is one of the most important causes of morbidity and mortality in the developed world and is gradually more frequent in the developing world including Saudi Arabia. According to the Saudi Cancer Registry report 2015, CRC is the most common cancer in men (14.9%) and the second most prevalent cancer. Oncogenic mutations in the KRAS gene play a central role in tumorigenesis and are mutated in 30–40% of all CRC patients. To explore the prevalence of KRAS gene mutations in the Saudi population, we collected 80 CRC tumor tissues and sequenced the KRAS gene using automated sequencing technologies. The chromatograms presented mutations in 26 patients (32.5%) in four different codons, that is, 12, 13, 17, and 31. Most of the mutations were identified in codon 12 in 16 patients (61.5% of all mutations). We identified a novel mutation c.51 G>A in codon 17, where serine was substituted by arginine (S17R) in four patients. We also identified a very rare mutation, c.91 G>A, in which glutamic acid was replaced by lysine (E31K) in three patients. In conclusion, our findings further the knowledge about KRAS mutations in different ethnic groups is indispensable to fully understand their role in the development and progression of CRC., GRAPHICAL ABSTRACT

Background Mutations in Kirsten rat sarcoma proto-oncogene (KRAS) have been shown to be associated with advanced-stage colorectal cancer (CRC), negative disease outcomes, and poor response to treatment. The purpose of this study was to investigate which CT features are biomarkers for KRAS gene mutation and impact the survival outcomes of colorectal cancer patients. Results Of the 113 CRC patients included in the study, 46 had KRAS mutations (40.71%) and 67 had no mutations (59.29%). Regional lymph node necrosis was the only imaging feature significantly associated with KRAS mutation (P = 0.011). Higher T staging and liver, lung, and distant metastasis were prognostic factors for CRC (P = 0.014, P < 0.001, P = 0.022, P < 0.001, respectively). There were no significant differences in overall survival between patients with KRAS mutations and those without (P = 0.159). However, in patients with no KRAS mutation, those with CRC on the left side had a significantly higher rate of survival than those with CRC on the right (P = 0.005). Conclusion Regional lymph node necrosis may be an imaging biomarker of CRC with KRAS mutation, possibly indicating poor prognosis.

Ovarian-like epithelial tumors of the testis, including serous borderline tumors, are rare entities. We report the case of a 60-year-old man with a left intratesticular mass who had a radical orchidectomy. Histologically, the tumor was identical to the ovarian counterpart showing a well-delineated cystic lesion characterized by intraluminal papillae. The papillae are lined by atypical cuboidal or ciliated cells and are associated with psammoma bodies. The tumor cells express cytokeratin 7 (CK7), cytokeratin 5-6 (CK5-6), cancer antigen 125 (CA125), estrogen (ER), progesterone (PR), Wilm’s tumor gene (WT1), paired box gene 8 (PAX8), Ber-EP4, and epithelial membrane antigen (EMA). The diagnosis of a serous borderline tumor of the testis was proposed. Mutation testing using next-generation sequencing showed a Q61K KRAS gene mutation. To the best of our knowledge, this is the second case report of a serous borderline tumor of the testis with a Q61K KRAS gene mutation.

Background: Colorectal cancer is the third most common cancer according to American Cancer Society. It is also the third most common cause of death in men and women in US. Colorectal cancer encompasses 5% of all cancer and 29% of gastrointestinal cancer with men and women ratio is about 3:1. More than 1/3 of colorectal cancer occur below the age of 45 years. Mutation in Kirsten rat sarcoma (KRAS) gene was found in 30-50% of colorectal cancer in which it was suggested to associated with increase proliferation and decrease apoptosis. This study aimed to analyze the association between diet profile and KRAS gene mutation.Method: This study was a cross sectional study. Data was collected from medical records of colorectal cancer patient in Dr. Saiful Anwar General Hospital, which included KRAS gene mutation analysis.Results: There were 12 subjetcs included in this study. Four subjects (33.3%) had gene mutation with 3 subjects (75%) had positive KRAS mutation and 1 subjects (25%) had positive Neuroblastoma rat sarcoma (NRAS) mutation. In non-mutation group, it had been found a higher frequency of green leaf vegetables diet, in comparison with mutation group (p = 0.023). There was positive correlation between green leaf vegetables diet with gene mutation.Conclusion: Mutation of KRAS and NRAS mutation in colorectal carcinoma were found in 33.3% of subjects. Data analysis showed positive association between low green leaf vegetables diet with KRAS and NRAS mutation.

In 2016 3158 patients with colorectal cancer (CRC) were registered in the Republic of Kazakhstan, out of them, 1,484 patients died. Colorectal and colon cancers are the 8th and the 5 th leading causes of mortality, respectively. One of the most significant events in the molecular pathogenesis of CRC is an activating mutation in the KRAS oncogene. Recently, studies of the KRAS gene mutation and analysis of its relation with the clinical course of CRC have been carried out in different countries. The effect of gender and age on the KRAS gene status in CRC remains a subject for discussion. The purpose of this study was to study the relationship between the KRAS gene status and gender, age and race in colorectal cancer patients residing in the Republic of Kazakhstan. Material and methods. Data on 332 patients with CRC for the period from 2010 to 2014 were studied. KRAS test was performed using BioLink kits to detect mutations in 12 and 13 codons of 2 exon using allele-specific PCR method. R esults. In our study, the frequency of KRAS gene mutations in CRC patients residing in the Republic of Kazakhstan was 44.9 %. The frequency of mutations in the studied codons among women and men was the same. When analyzing the mutation frequency of the KRAS gene, it was revealed that among both sexes the G12D mutation was more often observed. The least common mutations were G12C, G12S, which occurred in both sexes (up to 5 % of cases). The analysis of the dependence of the mutation on the race of the patients revealed some predominance of the wild type in the Asian group – 94 (51.4 %), while Europeans were more often detected with the KRAS mutation – in 81 (54.4 %) patients. Mutation of the KRAS gene was more frequently observed in the group of older patients.

Pancreatic ductal adenocarcinoma (PDAC) patients eligible for curative surgery undergo unpredictable disease relapse. Even patients with a good pathological response after neoadjuvant treatment (NAT) remain susceptible to recurrent PDAC. Molecular analysis of R0 margins may identify patients with a worse prognosis. The molecular status of mutant KRAS (exon 2, codon 12/13) was analysed retrospectively by digital droplet PCR in tumour areas, venous and resection margins of resected tumours, either undergoing up-front surgery (UFS) or after NAT with a good pathological response. Expectedly, tumour tissues or remnants from patients who underwent NAT presented lower KRAS mutant allele frequencies (MAF) than patients eligible for UFS. Similarly, ypT1 tumour MAFs were greater than the ypT0 tumour remnant MAFs in the NAT group. Mutant KRAS status in margins did not distinguish NAT subgroups. It was not predictive of shorter recurrence-free or overall survival within or between groups. KRAS-double negativity in both venous and resection margins did not identify patients with a better prognosis, regardless of the groups. The cohorts ‘sizes were small due to limited numbers of patients meeting the inclusion criteria, but KRAS-positivity or MAFs in resection and venous margins did not carry prognostic value. Comparison of margins from good versus bad responders receiving NAT may provide better clinical value.

Pancreatic carcinoma (PC) is greatly induced by the KRAS gene mutation, but effective targeted delivery for gene therapy has not existed. Small interfering ribonucleic acid (siRNA) serves as an advanced therapeutic modality and holds great promise for cancer treatment. However, the development of a non-toxic and high-efficiency carrier system to accurately deliver siRNA into cells for siRNA-targeted gene silencing is still a prodigious challenge. Herein, polyethylenimine (PEI)-modified hydroxyapatite (HAp) nanoparticles (HAp-PEI) were fabricated. The siRNA of the KRAS gene (siKras) was loaded onto the surface of HAp-PEI via electrostatic interaction between siRNA and PEI to design the functionalized HAp-PEI nanoparticle (HAp-PEI/siKras). The HAp-PEI/siKras was internalized into the human PC cells PANC-1 to achieve the maximum transfection efficiency for active tumor targeting. HAp-PEI/siKras effectively knocked down the expression of the KRAS gene and downregulated the expression of the Kras protein in vitro. Furthermore, the treatment with HAp-PEI/siKras resulted in greater anti-PC cells’ (PANC-1, BXPC-3, and CFPAC-1) efficacy in vitro. Additionally, the HAp-PEI exhibited no obvious in vitro cytotoxicity in normal pancreatic HPDE6-C7 cells. These findings provided a promising alternative for the therapeutic route of siRNA-targeted gene engineering for anti-pancreatic cancer therapy.

Objective: KRAS mutation occurs in 30% to 50% of colorectal cancers. The aim of our study was to determine the frequency of KRAS mutations among patients with colorectal cancer; and the relationship with clinicopathologic features. Materials and Methods: 79 colorectal cancer cases at a hospital in Hai Phong of Vietnam were collected, including 45 colon cancer and 34 rectal cancer during January 2010 and July 2012. PCR amplification and DNA sequencing were used to detect mutations in exon 2 of KRAS gene. The study was based on informed consent and approval by the Ethics Committee of Viet Tiep Hospital. Results: KRAS mutation was found in 40.4% (225/557) colorectal cancer. All mutation locations were in codon 12. There was significant association (p < 0.05) between KRAS mutations, tumor size and tumor stage. No significant association was observed between KRAS mutations and gender, tumor location, tumor grade or histologic presence of mucin (p>0.05). Conclusion: Determining the KRAS mutational status of tumor samples has become an essential tool for managing patients with colorectal cancers Keywords: colorectal cancer, KRAS gene mutation, clinicopathology.

To investigate the association between KRAS mutation and prognosis in rectal cancer patients with neoadjuvant chemoradiotherapy. Literature was searched in the databases including Cochrane Library, EMBASE (Ovid), and MEDLINE (PubMed) from inception to December 16, 2020. The keywords “rectal cancer” or “rectal carcinoma” or “rectal adenocarcinoma” and “KRAS” and “neoadjuvant” were used for preliminary literature retrieval. Pooled odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated for the KRAS mutation and clinical outcomes including overall survival (OS), disease-free survival (DFS), pathologic complete remission (pCR), downstaging of T stages and tumor stages, as well as improvements in tumor regression grading (TRG). Publication bias was assessed by the funnel plots. A total of 16 articles were included for eligibility. The total number of patients was 3674 cases, with an incidence of KRAS gene mutation of 36.6% (1346/3674). Meta-analysis showed that the pooled OR for KRAS mutation on OS was 1.33 (95%CI: 113–1.56). Consistently, results also indicated that the KRAS mutant was related to the poor DFS (pooled OR=1.55, 95%CI: 1.19–2.02). However, KRAS mutation is not related to the PCR (pooled OR= 0.71, 95%CI: 0.44–1.14), downstaging in T stages (pooled OR= 0.66, 95%CI: 0.42–1.06), tumor stages (pooled OR= 1.18, 95%CI: 0.78–1.78, I2=12.9%), as well as improvement in TRG grades (pooled OR= 0.84, 95%CI: 0.59–1.20). KRAS mutation is a predictor for the poor prognosis of neoadjuvant chemoradiotherapy in patients with rectal cancer, but it is not related to the responses of tumors after treatment.

Papillary renal neoplasm with reverse polarity (PRNRP) is a recently proposed entity of renal tumor. It shows a far better prognosis than papillary renal cell carcinoma (PRCC) and frequently has KRAS missense mutation. In this study, we compared 14 cases of PRNRP and 10 cases of PRCC type 1 (PRCC1) and type 2 (PRCC2) from clinical, morphological, immunohistochemical, and molecular biological perspectives. We subjected all PRNRP and PRCC cases to immunohistochemical analysis. Whole-exome sequencing using next-generation sequencing (NGS) was performed for six cases of PRNRP, three cases of PRCC1, and four cases of PRCC2. A search for KRAS gene mutation in the remaining eight cases of PRNRP was performed by polymerase chain reaction (PCR) sequencing. The results showed that all cases of PRNRP were pT1N0M0, none of which followed a course of recurrence or tumor-related death. Immunohistochemical analysis revealed diffuse staining of CK7, EMA, PAX8, and GATA3 but weak or negative staining of CD10, CD15, and AMACR in PRNRP. By NGS and PCR, KRAS missense mutation was detected in 11 of 14 PRNRP cases, although pathogenic KRAS mutation was not observed in PRCC1 and PRCC2. NGS analysis revealed less tumor mutation burden in PRNRP than in PRCC. PRNRP also showed no specific chromosomal copy number abnormalities, including gains of 7 and 17. In conclusion, we propose that PRNRP is a distinct condition from PRCC.

Background Mutations in the RAS genes, HRAS, KRAS, and NRAS, are the most common modifications in many types of human tumors and are found in approximately 30% of all human cancers. These mutations are usually found in codons 12, 13, or 61. Methods The aim of this study is to evaluate mutations in codons 59, 117, and 146 of KRAS and NRAS genes in addition to codons 12,13, and 61 of KRAS gene in lung cancer tissue specimens obtained with bronchoscopy. KRAS and NRAS mutation analyses with pyrosequencing were performed on DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples of 64 patients histopathologically diagnosed as lung cancer after bronchoscopic biopsy. Results In all, 20 patients (31.2%) had mutations in KRAS gene (8/27 squamous cell carcinoma, 8/11 adenocarcinoma, 3/16 small cell carcinoma, and 1/1 pleomorphic carcinoma). The most common mutation in codon 12 was in c.35G>T (G12V). When the mutation rate of adenocarcinoma (72.7%) and squamous cell carcinoma (22.9%) patients was compared with each other, a statistically significant difference was observed (P = 0.008). There were no mutations in codons 59, 117, or 146 of KRAS and NRAS genes in patients with lung cancer. Conclusion In this study, we firstly examined mutations in codons 59, 117, and 146 of KRAS and NRAS genes in addition to codons 12, 13, and 61 of KRAS gene in Turkish lung cancer patients both in non-small cell lung cancer and small cell lung cancer. Although no mutation was detected in codons 59, 117, and 146 of KRAS and NRAS genes, the frequency of KRAS gene mutation was higher than the rate of mutation in both Asian and Western countries, and multicenter studies including more cases should be performed to further explore our results.

Objective. The main objective of the study was to determine the effect of the presence of mutation in the KRAS gene on the survival in patients with colorectal cancer (CRC) and peritoneal metastases (PM). Materials and Methods. A retrospective cohort study was performed. Patients diagnosed with CRC with synchronous or metachronous PM between January 2006 and December 2019 were included. Data on the histopathological, clinical, and treatment factors were collected. The effect of each variable on survival was evaluated by Cox regression. Results. A total of 149 patients were included (64 women (43%) and 85 men (57%); mean age, 63 years). The long-term survival rate at 36 months was 24% (median, 21 months). KRAS mutation was detected in 75 patients (50.3%). Kaplan–Meier analysis estimated that likelihood of survival was higher in patients with wild-type KRAS tumours (35%) than in mutated-type KRAS (14%) (median: 28 vs. 15, respectively) ( P = 0.001 ). Within the categories into which the peritoneal cancer index (PCI) was classified, survival at 36 months depended on the KRAS status. Survival in wild-type KRAS tumours with PCI 1–10 was 71% and with PCI 11–20 was 26%, while in mutant-type KRAS tumours, survival was 41% and 4%, respectively ( P = 0.025 ). In the multiple regression analysis, the KRAS mutation was revealed to have an independent prognostic value (HR: 2.144; 95% CI: 1.342–3.424). Conclusion. The mutational status of the KRAS gene has demonstrated a strong association with survival and prognostic utility in patients with CRC with PM.

BACKGROUND: Raise of ovarian cancer mortality is caused by high ovarian cancer recurrence. This is related to many prognostic factors. Kirsten-rat sarcoma virus oncogene (KRAS) is a proto-oncogene that regulates proliferation, growth and cell motility. The controversy of some experts regarding KRAS mutations in the prognosis of ovarian cancer makes it interesting to analyze. AIM: The aim of this study is to clarify whether the clinicopathologic factors and KRAS gene mutation affect the recurrence of patients with ovarian cancer in Indonesia. METHODS: The authors conducted a retrospective cohort study. Clinicopathological factors and prognoses were obtained for 205 patients who were histopathologically diagnosed with epithelial ovarian cancer or ovarian borderline malignant tumor, operated from June 2015 to January 2019 at Dr. M. Djamil General Hospital. We gathered 80 patients who were diagnosed with epithelial ovarian cancer since June 2015 until January 2019. These cases were analyzed after 2-year follow-up or recurrence occurred. Survival rate was determined using the Kaplan–Meier method and examined by Log rank test. All analyses were performed using STATA ver. 12.0, with p < 0.05 considered to be significant. RESULTS: Among KRAS mutation group, the 2-year disease free survival rate (2y-DFS) was 31.56% and 47.58% in non-mutation group with significant differences between mutation and non-mutation (p = 0.02). There was a significant difference between early stage ovarian cancer with non-mutation group and advanced stages ovarian cancer with mutation group (p = 0.00). Among combination staging with mutation group, the 2y-DFS was 85.79% in early stage ovarian cancer with non-mutation, 44.44% in early stage with mutation, 10.65% in advanced stage with non-mutation, and 20.00% in advanced stage with mutation. CONCLUSION: The results suggest that staging and KRAS mutation are the most influence prognostic factors for epithelial ovarian cancer. There was a discrepancy of prognosis by staging and mutation between early stage with non-mutation and advanced stage with KRAS mutation.

Background The presence of histologically different neoplasms in the same organ is rare in pathologic practice. We report the first case of synchronous clear cell renal cell carcinoma (clear cell RCC) and papillary renal neoplasm with reverse polarity (PRNRP) with comprehensive immunohistochemical and molecular characterization using next-generation sequencing (NGS). Case presentation A 61-year-old man was incidentally found to have a left renal mass on imaging studies performed for workup of left back pain and urine color change for 1 week. A laparoscopic left radical nephrectomy was performed. Gross examination showed lobulated masses measuring 5.6 × 4.0 × 3.3 cm in the upper to mid pole and 1.1 × 1.0 × 1.0 cm in the lower pole. Microscopic findings revealed these to be two different separate masses of clear cell renal cell carcinoma and papillary renal neoplasm with reverse polarity. NGS analyses revealed KRAS gene mutation (c.35G > T/p.G12V in exon 2) in the papillary renal neoplasm with reverse polarity, with PIK3CA gene mutation restricted to the clear cell renal cell carcinoma (c.1624G > A/p.E542K in exon 10). Conclusions We report here an extraordinarily rare case of synchronous renal tumors of papillary renal neoplasm with reverse polarity and clear cell renal cell carcinoma. We identified simultaneous KRAS and PIK3CA mutations in two different renal masses in the same kidney for the first time. New pathologic assessment with comparative molecular analysis of mutational profiles may be helpful for tumor studies.

espanolRESUMEN Objetivos: el objetivo principal del estudio fue evaluar la supervivencia global en pacientes con carcinoma colorrectal metastasico (CCRm) cuyos tumores tuvieran el gen KRAS mutado frente al no mutado. Materiales y metodos: se analizaron los datos de las historias clinicas de pacientes con CCRm (enero 2010 - diciembre 2013) de diferentes hospitales de Lima Metropolitana, cuyos tumores tuvieron evaluacion del estado mutacional del exon 2, gen KRAS. Se usaron la curva de supervivencia de Kaplan-Meier y la prueba de long rank o Breslow para las comparaciones de las curvas de supervivencia. Resultados: de los 320 casos analizados, hubo 227 pacientes (70,93%) con KRAS no mutado y 93 (29,07%) con KRAS mutado. La supervivencia global de pacientes con CCRm y KRAS mutado fue mayor que los pacientes con KRAS no mutado (hazart ratio: 0,73; IC 95% 0,55 - 0,98; p=0,037). Conclusion: la poblacion con CCRm y KRAS mutado estudiada en centros medicos de Lima Metropolitana tuvo una supervivencia mayor, comparada con la no mutada, comportamiento diferente a lo encontrado en la literatura mundial. EnglishABSTRACT Objectives : The main goal for this study was to evaluate overall survival in mCRC patients with mutated vs. wild type KRAS gene (exon 2) status. Materials and Methods : Between January 2010 and December 2013, data from clinical records of mCRC patients from different hospitals in Lima, stating an assessment of the KRAS gene mutation status (exon 2), were analyzed. Kaplan-Meier survival estimates and Log-Rank or Breslow Test were used to compare the survival curves. Results : Three-hundred and twenty cases were analyzed. There were 227 patients (70.93%) with wild type KRAS and 93 (29.07%) with mutated KRAS. The overall survival of mCRC patients and mutated KRAS was higher than that of patients with wild type KRAS (HR: 0.73; 95% CI: 0.55-0.98; P= 0.037). Conclusion : Patients with mCRC and mutated KRAS who were studied in Lima have higher survival rates compared to wild type patients, being this different from what is found in the world literature.

To investigate the association between metabolic parameters of dual time point 18F-FDG PET/CT imaging and Kirsten rat sarcoma (KRAS) mutation status in colorectal liver metastases (CRLM). Forty-nine colorectal cancer patients with 87 liver metastatic lesions were included in this retrospective study. KRAS gene mutation tests were also performed for all the patients. The maximum standardized uptake value (SUVmax) was measured for each hepatic metastatic lesion on both early and delayed scans, and the change of SUVmax (ΔSUVmax) and retention index (RI) were calculated. Uni-variate and multi-variate analyses were employed to determine the relationship between any PET/CT parameters and KRAS mutation status. Thirty-seven (42.5%) liver metastatic lesions harboring KRAS mutations were identified. The SUVmax of CRLM with KRAS mutation both on early and delayed scans was significantly higher than those with wild-type KRAS (10.7 ± 6.0 vs. 7.8 ± 3.3, P = 0.002; 15.5 ± 10.1 vs. 10.0 ± 4.2, P

Objective: to assess the efficacy of anti-angiogenic agents incorporated into second-line chemotherapeutic regimens for metastatic colon cancer depending on the KRAS gene mutation status.Materials and methods. We selected completed prospective randomized controlled phase III clinical trials evaluating the efficacy of antiangiogenic agents (bevacizumab, ramucirumab and aflibercept) added to second-line chemotherapy for metastatic colon cancer with subanalysis of treatment efficacy depending on the KRAS gene mutation status. Meta-analysis was performed using the ReviewManager (RevMan) v. 5.3 (The Cochrane Collaboration, Denmark).Results. Three studies (ML18147, RAISE, VELOUR) involving 2165 patients (1137 with KRAS wild-type tumors and 1028 with KRAS-mutant tumors) met the inclusion criteria and were included into this meta-analysis. The majority of patients (84 %) received bevacizumab in the first-line treatment. The results of our meta-analysis suggest that adding an anti-angiogenic drug to chemotherapy in patients with KRAS wildtype colon cancer significantly improved both progression-free survival (hazard ratio (HR) 0.71; 95 % confidence interval (CI) 0.62–0.80; р= 0.0001; I2 = 0, p = 0.59). In patients with KRASmutant colon cancer, incorporation of an anti-angiogenic drug into the treatment regimen was not associated with better overall survival (ОР0.9; 95 % CI 0.79–1.03; р= 0.11; I2 = 0, p = 0.98), although improved progression-free survival (HR 0.78; 95 % CI 0.68–0.89; р= 0.0002; I2 = 0, p = 0.46). Conclusion. Continuation of anti-angiogenic therapy in the second-line treatment for metastatic colon cancer is most effective in patients with KRAS wild-type tumors. In individuals with KRAS-mutant tumors, continuation of bevacizumab or switch to another anti-angiogenic agent in the second-line treatment improves progression-free survival and has a statistically insignificant effect on overall survival.

// Ryogo Minamimoto 1 , Mehran Jamali 1 , Olivier Gevaert 1 , Sebastian Echegaray 1 , Amanda Khuong 2 , Chuong D. Hoang 2 , Joseph B. Shrager 2 , Sylvia K. Plevritis 1 , Daniel L. Rubin 1 , Ann N. Leung 1 , Sandy Napel 1 and Andrew Quon 1 1 Department of Radiology, Stanford University, Stanford, CA, USA 2 Department of Cardiothoracic Surgery, Stanford University, Stanford, CA, USA Correspondence to: Andrew Quon, email: aquon@stanford.edu Keywords: 18 FDG-PET/CT, heterogeneity, KRAS gene mutation, EGFR gene mutation, NSCLC Received: September 16, 2016 Accepted: March 20, 2017 Published: May 10, 2017 ABSTRACT This study investigated the relationship between epidermal growth factor receptor ( EGFR ) and Kirsten rat sarcoma viral oncogene homolog ( KRAS ) mutations in non-small-cell lung cancer (NSCLC) and quantitative FDG-PET/CT parameters including tumor heterogeneity. 131 patients with NSCLC underwent staging FDG-PET/CT followed by tumor resection and histopathological analysis that included testing for the EGFR and KRAS gene mutations. Patient and lesion characteristics, including smoking habits and FDG uptake parameters, were correlated to each gene mutation. Never-smoker ( P < 0.001) or low pack-year smoking history ( p = 0.002) and female gender ( p = 0.047) were predictive factors for the presence of the EGFR mutations. Being a current or former smoker was a predictive factor for the KRAS mutations ( p = 0.018). The maximum standardized uptake value (SUV max ) of FDG uptake in lung lesions was a predictive factor of the EGFR mutations ( p = 0.029), while metabolic tumor volume and total lesion glycolysis were not predictive. Amongst several tumor heterogeneity metrics included in our analysis, inverse coefficient of variation (1/COV) was a predictive factor ( p < 0.02) of EGFR mutations status, independent of metabolic tumor diameter. Multivariate analysis showed that being a never-smoker was the most significant factor ( p < 0.001) for the EGFR mutations in lung cancer overall. The tumor heterogeneity metric 1/COV and SUV max were both predictive for the EGFR mutations in NSCLC in a univariate analysis. Overall, smoking status was the most significant factor for the presence of the EGFR and KRAS mutations in lung cancer.

The KRAS gene mutation is involved in several types of tumors. However, the potential role of the KRAS mutation in human primary and paired metastatic colorectal cancer (CRC) among different nationalities is poorly understood. In the present study, we assessed the relationship between KRAS mutation status and overall survival (OS) and disease-free survival (DFS) in 230 patients with primary and paired metastatic CRC. The KRAS mutation rate in primary CRC tissue was 43.0% (99/230), which was higher than in paired metastatic CRC, which was 31.9% (23/72; P

KRAS gene mutation causes lung adenocarcinoma. KRAS activation has been associated with altered glucose and glutamine metabolism. Here, we show that KRAS activates lipogenesis, and this activation results in distinct proteomic and lipid signatures. By gene expression analysis, KRAS is shown to be associated with a lipogenesis gene signature and specific induction of fatty acid synthase (FASN). Through desorption electrospray ionization MS imaging (DESI-MSI), specific changes in lipogenesis and specific lipids are identified. By the nanoimmunoassay (NIA), KRAS is found to activate the protein ERK2, whereas ERK1 activation is found in non-KRAS-associated human lung tumors. The inhibition of FASN by cerulenin, a small molecule antibiotic, blocked cellular proliferation of KRAS-associated lung cancer cells. Hence, KRAS is associated with activation of ERK2, induction of FASN, and promotion of lipogenesis. FASN may be a unique target for KRAS-associated lung adenocarcinoma remediation.

Key Clinical Message While the role of KRAS gene mutations has been widely accepted for predicting responses to anti‐EGFR therapy in patients with colorectal cancer, although this study was based on observation of a single case it gives hope that some KRAS gene mutation may have favorable prognosis. More studies are required on patients with similar mutation to validate this finding.

Tissue damage increases the risk of cancer through poorly understood mechanisms1. In mouse models of pancreatic cancer, pancreatitis associated with tissue injury collaborates with activating mutations in the Kras oncogene to markedly accelerate the formation of early neoplastic lesions and, ultimately, adenocarcinoma2,3. Here, by integrating genomics, single-cell chromatin assays and spatiotemporally controlled functional perturbations in autochthonous mouse models, we show that the combination of Kras mutation and tissue damage promotes a unique chromatin state in the pancreatic epithelium that distinguishes neoplastic transformation from normal regeneration and is selected for throughout malignant evolution. This cancer-associated epigenetic state emerges within 48 hours of pancreatic injury, and involves an ‘acinar-to-neoplasia’ chromatin switch that contributes to the early dysregulation of genes that define human pancreatic cancer. Among the factors that are most rapidly activated after tissue damage in the pre-malignant pancreatic epithelium is the alarmin cytokine interleukin 33, which recapitulates the effects of injury in cooperating with mutant Kras to unleash the epigenetic remodelling program of early neoplasia and neoplastic transformation. Collectively, our study demonstrates how gene–environment interactions can rapidly produce gene-regulatory programs that dictate early neoplastic commitment, and provides a molecular framework for understanding the interplay between genetic and environmental cues in the initiation of cancer. In mouse models of pancreatic cancer, a cooperative interaction between tissue damage and Kras gene mutation rapidly induces cancer-associated chromatin states in pre-malignant tissue, leading to gene dysregulation and neoplastic transformation.

The colorectal cancer is considered as the third most common malignant disease in the world. During last decade, the problem of colorectal cancer became especially urgent conditioned not only by increasing of number of metastatic tumors of colon and rectum, but also by implementation of high-tech methods of treatment that significantly improved the results of five-year survival period. Along with success of therapy, understanding of genomics of colorectal cancer and owing to extensive application of next generation sequencing, the opportunity of optimal choice of treatment options was offered.Methods. The sampling included tumor material of 332 patients diagnosed with colorectal cancer, fixed in formalin and enclosed in paraffin, and treated in oncology dispensaries and centers and in the Kazakh Research Institute of Oncology and Radiology in 2010-2014. After morphological evaluation of the material quality, the molecular genetic analysis was applied to establish mutation of the KRAS gene using polymerase chain reaction in real-time.Results. The study established that KRAS mutation rate in patients with colorectal cancer had no reliable dependency with the region of their residence, since statistical analysis of pair “region of residence - all KRAS mutations” had value of correlation coefficient rp = 0.1 (p = 0.05).Conclusions. colorectal cancer It was established that there is no reliable dependence of rate of KRAS gene mutation in patients with colorectal cancer in the region of their residence. The high rate of occurrence of mutated type of gene was detected in 3 regions (Kyzyl-Orda, North Kazakhstan and Almaty) within the range of 58.3-76.5% of cases. The wild type of KRAS gene was most frequently detected in patients from the Karaganda region (65.5%), South Kazakhstan (71.4%) and Almaty (60.7%).

A 12-year-old female patient with cardiofaciocutaneous syndrome in the presence of a KRAS gene mutation had episodes of pericardial effusion on ultrasound, later confirmed to be chylopericardium, which resolved after a lymphangiography. We discussed herein the pathophysiological background of this rare case and the efficacy of lymphangiography in the treatment of chylopericardium.

Introduction: Colorectal cancer is currently the third most common cancer in terms of incidence and mortality in Iran. Different gene mutations may confer different degrees of biological aggressiveness and reduce the effectiveness of targeted therapeutic strategies in metastatic Colorectal Cancer (mCRC) patients. Aim: To evaluate any mutation pattern in mCRC, and then evaluate clinical and epidemiological correlations with the detected mutations. Materials and Methods: This study was a cross-sectional analytical study, and all mCRC cases referred to two main central hospitals and oncologists’ clinics, in Tabriz, Iran, from January 2016 to November 2018 were enrolled. KRAS, NRAS, and BRAF mutation tests were performed routinely for all mCRC cases before considering any treatment strategies. Idylla Biocartis NV system{Test Type Package (TTP)}, determined the presence of mentioned mutations. Logistic regression models were used to statistically analysis. Results: The present authors included 173 cases with confirmed mCRC. Among 102 patients with KRAS gene mutation detection, the frequency of mutations was 38.23% (n=39) while most were in exon 2, codon 12 (61.54%), followed by patients who had mutations in codon 13 (n=5, 12.82%). NRAS mutations were only observed in one patient (1.33%) from among the 75 cases who were tested. BRAF codon 600 was tested in 39 cases, and only one case (2.56%) had the mutation. Patients with left-sided tumours had about 9.5 times higher likelihood of KRAS mutation than right-sided tumours (OR=9.64; 95% CI=1.24-75.27). Smoking increased the odds of KRAS mutation about 50%, and mCRC who had alcohol consumption had about two times more likelihood of mutation. Conclusion: The overall frequency of KRAS mutations in the present study was high, while the frequency of KRAS mutations in mCRC patients is lower in Asian populations. KRAS mutation results in this study were most similar to European populations.

KRAS is a member of the murine sarcoma virus oncogene-RAS gene family. It plays an important role in the prevention, diagnosis and treatment of tumors during tumor cell growth and angiogenesis. KRAS is the most commonly mutated oncogene in human cancers, such as pancreatic cancers, colon cancers, and lung cancers. Detection of KRAS gene mutation is an important indicator for tracking the status of oncogenes, highlighting the developmental prognosis of various cancers, and the efficacy of radiotherapy and chemotherapy. However, the efficacy of different patients in clinical treatment is not the same. Since RNA interference (RNAi) technologies can specifically eliminate the expression of specific genes, these technologies have been widely used in the field of gene therapy for exploring gene function, infectious diseases and malignant tumors. RNAi refers to the phenomenon of highly specific degradation of homologous mRNA induced by double-stranded RNA (dsRNA), which is highly conserved during evolution. There are three classical RNAi technologies, including siRNA, shRNA and CRISPR-Cas9 system, and a novel synthetic lethal interaction that selectively targets KRAS mutant cancers. Therefore, the implementation of individualized targeted drug therapy has become the best choice for doctors and patients. Thus, this review focuses on the current status, future perspective and associated challenges in silencing of KRAS with RNAi technology.

Simple Summary The vast majority of patients with pancreatic ductal adenocarcinomas harbor KRAS mutations in their tumors. Functionally, mutated KRAS is not only dedicated to tumor cell proliferation, survival and invasiveness, but also causing the immunosuppression in this cancer. In this situation, current data indicating the therapeutic effects of immune checkpoint inhibitors on pancreatic ductal adenocarcinomas are still not satisfying. In order to reflect the present bottleneck of immune checkpoint inhibitors in managing this cancer, we mainly provide information associated with the mechanism by which KRAS mutations establish the immunosuppressive milieus in pancreatic ductal adenocarcinomas. Together with other advances in this field, future directions to overcome the KRAS mutation-induced immunosuppression in pancreatic ductal adenocarcinomas are raised as well. Meanwhile, lung adenocarcinomas and colorectal adenocarcinomas are enumerated to compare with pancreatic ductal adenocarcinomas, aiming to indicate the specificity of KRAS mutations in dictating tumoral immune milieus among these cancers. Abstract Generally, patients with pancreatic ductal adenocarcinoma, especially those with wide metastatic lesions, have a poor prognosis. Recently, a breakthrough in improving their survival has been achieved by using first-line chemotherapy, such as gemcitabine plus nab-paclitaxel or oxaliplatin plus irinotecan plus 5-fluorouracil plus calcium folinate. Unfortunately, regimens with high effectiveness are still absent in second- or later-line settings. In addition, although immunotherapy using checkpoint inhibitors definitively represents a novel method for metastatic cancers, monotherapy using checkpoint inhibitors is almost completely ineffective for pancreatic ductal adenocarcinomas largely due to the suppressive immune milieu in such tumors. Critically, the genomic alteration pattern is believed to impact cancer immune environment. Surprisingly, KRAS gene mutation is found in almost all pancreatic ductal adenocarcinomas. Moreover, KRAS mutation is indispensable for pancreatic carcinogenesis. On these bases, a relationship likely exists between this oncogene and immunosuppression in this cancer. During pancreatic carcinogenesis, KRAS mutation-driven events, such as metabolic reprogramming, cell autophagy, and persistent activation of the yes-associated protein pathway, converge to cause immune evasion. However, intriguingly, KRAS mutation can dictate a different immune environment in other types of adenocarcinoma, such as colorectal adenocarcinoma and lung adenocarcinoma. Overall, the KRAS mutation can drive an immunosuppression in pancreatic ductal adenocarcinomas or in colorectal carcinomas, but this mechanism is not true in KRAS-mutant lung adenocarcinomas, especially in the presence of TP53 inactivation. As a result, the response of these adenocarcinomas to checkpoint inhibitors will vary.

Introduction. The concordance of KRAS gene mutation status between the primary and metastatic CRC is 95%. The aim of this study was to find factors associated with the disconcordance of KRAS, NRAS, BRAF, PIK3CA mutation status between the primary tumors and metastases in pts with CRC. Patients and methods. We performed DNA melting analysis with TaqMan probes and following Sanger sequencing to detect mutation hot-spots in KRAS exons 2 and 3, NRAS exons 2 and 3, BRAF exon 15, PIK3CA exons 9 and 20 in 148 tumor tissues from 65 pts (65 primary tumors and 83 metastases). Results. Mutations in KRAS, NRAS, PIK3CA and BRAF genes in primary tumors were detected in 43.1%, 3.1%, 13.8% and 3.1%, respectively. Discordance of mutation status of genes was identified in 29.2% of patients: 16.9% in KRAS, 3% in NRAS, 12.3% in PIK3CA and 3% BRAF status. In all cases of metastases in the brain we found the discordance in KRAS and PIK3CA mutation status (p=0.08). Also, peritoneal metastases had discordance in KRAS status (p=0.02). With the increase of period from removal of the primary tumor and metastases, the incidence rate of changes in the mutational status of the genes also increased. Conclusion. discordance of the mutational status of genes, especially in the long course of the disease, raises the question of repeating biopsies in the progression of the disease

// Joo Kyung Park 1 , Yoon Jung Lee 1 , Jong Kyun Lee 1 , Kyu Taek Lee 1 , Yoon-La Choi 2 , Kwang Hyuck Lee 1, 3 1 Division of Gastroenterology, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea 2 Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea 3 Department of Health Sciences and Technology, Samsung Advanced Institute for Health Sciences & Technology, Sungkyunkwan University, Seoul, South Korea Correspondence to: Kwang Hyuck Lee, email: lkhyuck@gmail.com Keywords: KRAS gene mutation, endoscopic ultrasound-guided fine needle aspiration, pancreatic ductal adenocarcinoma Received: December 19, 2015 Accepted: October 26, 2016 Published: December 10, 2016 ABSTRACT EUS-FNA becomes one of the most important diagnostic modalities for PDACs. However, acquired tissue specimens were sometimes insufficient to make a definite cytological diagnosis. On the other hand, KRAS mutation is the most frequently acquired genetic alteration found more than 90% of PDACs. To investigate the way to improve diagnostic accuracy for PDACs using both cytological examination and KRAS mutation analysis would be a great help. Therefore, the aims of this study were to evaluate usefulness of conventional cytological examination combined with KRAS mutation analysis with modified PCR technology to improve the sensitivity and the accuracy. We enrolled 43 patients with solid pancreatic masses and 86 EUS-FNA specimens were obtained. During the EUS-FNA, the needle catheter was flushed with 2 cc of saline and the washed fluid was collected for KRAS mutation analysis for the first 2 passes; PNAClamp™ KRAS Mutation Detection Kit. There were 46 specimens from the 23 PDACs and 40 specimens from the 20 other pancreatic diseases. The sensitivity, specificity and accuracy were as follows; conventional cytopathologic examination: 63%, 100% and 80%; combination of cytopathologic examination and K-ras mutation analysis: 87%, 100% and 93%. Furthermore, KRAS mutation was detected 11 out of 17 PDAC samples whose cytopathology results were inconclusive. KRAS mutation analysis with PNAClamp™ technique using washing fluid from EUS-FNA along with cytological examination may not only improve the diagnostic accuracy of PDACs, but also establish the platform using genetic analysis which would be helpful as diagnostic modality for PDACs.