Search

Although drug combination has proved to be an efficient strategy for clinic gastric cancer therapy, how to further improve their bioavailability and reduce the side effects are still challenges due to the low solubility and untargeted ability of drugs. Recently, newly emerging nanotechnology has provided an alternative for constructing new drug delivery systems with high targeting ability and solubility. In this study, a pH-responsive liposome (Liposome-PEO, LP) loaded with apatinib (AP) and cinobufagin (CS-1) was used for combinational therapy against gastric cancer after coating with a hybrid membrane (R/C). The results indicated that the constructed nanocomplex LP-R/C@AC not only efficiently killed tumor cells in vitro by inducing apoptosis, autophagy, and pyroptosis, but also significantly inhibited tumor invasion and metastasis via the VEGFR2/STAT3 pathway. Moreover, it showed stronger anti-tumor activity in gastric cancer-bearing mouse models, as compared to the sole drugs. A naturally-derived hybrid cell membrane coating bestowed nanocomplexes with enhanced biointerfacing including prolonged circulation time and targeting ability.

As a breast cancer subtype with high mortality in women, the efficient treatment of Triple-negative breast cancer (TNBC) is still a challenge due to its unique metastatic mode and poor prognosis. In this study, we developed a biomimetic nanodelivery system (denoted as GTDC@M-R NPs) based on erythrocyte membrane (M)-camouflaged graphene oxide quantum dots (GOQDs, G) for TNBC therapy. The TAT (T) and RGD (R) peptides were used to endow targeting accumulation ability of Gamabufotalin (CS-6, C) and doxorubicin (DOX, D) in tumor tissue. In vitro assay indicated good biocompatibility, prolonged blood circulation time (3-fold longer than GT NPs), and effectively enhanced cell and nucleus targeting capability of this nanosystem. Fluorescence activated cell sorter (FACS) analysis indicated that the combination of DOX and CS-6 induced TNBC cell apoptosis more than 89 % under the ratio of 10:1. In vivo assay indicated that the accumulation of GTDC@M-R NPs in tumor sites increased about 2-fold compared to naked GTDC NPs, which was accompanied by high tumor apoptosis rates through blocking chemotherapy-activated cyclooxygenase-2 (COX-2) and enhancing DOX's anti-tumor activity of chemical drugs (85%). Moreover, comparing with the control, the average number of lung metastatic nodules in tumor-bearing mice reduced 84%, the molecular mechanism of which is related to the down expression of COX-2, matrix metalloproteinase 9 (MMP9) and vascular endothelial growth factor (VEGF). Taken together, our results proved that the developed GTDC@M-R NPs can inhibit the growth and suppress metastasis of TNBC, which broaden our insights into the application of combinational strategy for efficient TNBC therapy. STATEMENT OF SIGNIFICANCE: In this study, we developed a biomimetic nanodelivery system (denoted as GTDC@M-R NPs) based on erythrocyte membrane (M)-camouflaged graphene oxide quantum dots (GOQDs, G) for TNBC therapy. The TAT (T) and RGD (R) peptides were used to endow targeting accumulation ability of Gamabufotalin (CS-6, C) and doxorubicin (DOX, D) in tumor tissue. These GTDC@M-R NPs indicated synergistic chemotherapy against TNBC cells through the precise cell and nuclear targeting, immune escape, and improved DOX sensitivity. A effective inhibition of tumor growth and metastasis was achieved by inhibiting Bcl-2/BAX, COX-2 and VEGF related signal pathways. Our finding suggests that the developed GTDC@M-R NPs present great treating effects in the preclinical models of TNBC, which broaden our insights into the application of combinational strategy for efficient TNBC therapy.

Purpose Metastasis is a crucial cause of the high mortality in patients with lung cancer. Long non-coding RNAs (lncRNAs) are emerging as important players in the development and progression of human cancers. Here, we aimed to identify metastasis-associated lncRNA and to study its roles in the migration and invasion of lung cancer cells. Materials and methods We screened differentially expressed lncRNAs between high- and low-metastatic lung cancer cell lines by using microarray and identified the target lncRNA TM4SF1-AS1. The effect of the TM4SF1-AS1 on the invasion and migration was evaluated through the wound healing experiment and transwell assay. The expression of related genes was assessed by RNA sequence and Western blotting. Results TM4SF1-AS1 was highly expressed in high metastatic lung cancer cell line, and it was also significantly up-regulated in lymph node metastatic lung cancer and was associated with lymph node metastasis. Overexpression of TM4SF1-AS1 promoted the migration and invasion of lung cancer cells. Overexpression of TM4SF1-AS1 decreased the expression of E-Cadherin and increased the expression of Vimentin, Snail and Twist, while knockdown of TM4SF1-AS1 exhibited the opposite trend. Furthermore, RNA sequence analysis revealed that some signaling pathways, including PI3K/AKT signaling pathway, were enriched upon TM4SF1-AS1 overexpression. Western blotting further confirmed that the PI3K/AKT signaling pathway was activated by TM4SF1-AS1. Conclusion This study illustrates that TM4SF1-AS1 promotes the migration and invasion of lung cancer cells by activating the PI3K/AKT signaling pathway. TM4SF1-AS1 might be a novel target of molecular treatment for lung cancer.

Background Epidermal growth factor-containing fibulin-like extracellular matrix protein 2 (EFEMP2), also known as fibulin-4, MBP1 and UPH1, is an extracellular matrix protein associated with a variety of tumors. The purpose of this study was to investigate the prognostic value and the function of EFEMP2 in lung cancer. Methods The mRNA and protein expression of EFEMP2 in lung normal and cancer tissues, lung cancer cell lines (A549, H460, H1299 and H1650) and normal epithelial cell line BEAS-2B were evaluated by immunohistochemistry, RT-qPCR and Western blotting. The Public databases (Oncomine and Kaplan-Meier plotter) were used to investigate the prognostic value of EFEMP2 in lung cancer. RNA interference (RNAi) and overexpression transfection were performed to detect the effects of EFEMP2 up- or down-regulation on lung normal and cancer cell proliferation, invasion and metastasis in vitro and in vivo. Results EFEMP2 was lowly expressed in lung cancer tissues and cells, and its low expression was associated with malignant phenotype and poor prognosis of lung cancer. The same conclusion had been drawn from the Public databases. EFEMP2 overexpression significantly inhibited the invasion of lung cancer cells, hampered the process of EMT, and decreased the expression and activity of MMP2 and MMP9, while EFEMP2 knockdown remarkably enhanced the invasion of lung cancer cells, promoted EMT, and increased the expression and activity of MMP2 and MMP9. Conclusion The low expression of EFEMP2 was detected in lung cancer and was positively correlated with the poor prognosis of patients. EFEMP2 was a tumor suppressor gene that inhibited the progress of lung cancer, which suggested a new research objective for the future studies.

Runt‑related transcription factor 3 (RUNX3) is a candidate tumor suppressor, and its inactivation may play a crucial role in the carcinogenesis process of numerous cancer types, including esophageal squamous cell carcinoma (ESCC). We previously revealed that RUNX3 inactivation was correlated with lymph node metastasis (LNM) and ESCC recurrence. However, the exact mechanisms of this process are still under investigation. The aim of the present study was to examine the potential roles and underlying molecular mechanisms of RUNX3 in ESCC metastasis and the epithelial‑mesenchymal transition (EMT). According to the results, RUNX3 expression in ESCC tissue was significantly reduced compared with that in adjacent normal tissue (0.50±0.20 vs. 0.83±0.16; P

Signal transduction and activators of transcription factor (STAT) 3 is associated with a poor prognosis in certain types of cancer. The purpose of the present study was to investigate the clinical and prognostic significance of STAT3/p-STAT3 expression in esophageal squamous cell cancer (ESCC) patients. A total of 71 patients were enrolled in the study. STAT3 and p-STAT3 expression were detected by Western Blot and immunohistochemistry assays. Stattic, the STAT3 inhibitor, was used to block the activation of STAT3 in ESCC cell lines Eca-109 and Kyse-30, and the CCK8 assay was performed to detect the effect of Stattic on the viability of ESCC cells. The expression of associated genes was assessed by RT-PCR and Western blot at RNA and protein levels, respectively. STAT3 expression was correlated with infiltration degree (pT) and pTNM. And p-STAT3 expression was correlated with pT, lymphatic metastasis (pN) and pTNM. The expression of VEGF, Bcl-xl and Cyclin D1 was up-regulated in ESCC tissues and positively correlated with p-STAT3 level, besides Bcl-xl. In vitro, Stattic inhibited the viability of Eca-109 and Kyse-30 cells in a dose- and time- dependent manner, and significantly inhibited the expression of VEGF, Bcl-xl and CyclinD1 at mRNA and protein level. The 5-year survival rate of the 71 patients was significantly associated with pT, pN, pTNM stage, p-STAT3 level, VEGF expression and Cyclin D1 expression. pN and p-STAT3 expression were independent relevant factors. Our results showed that p-STAT3 might serve as an essential biomarker for tumor invasion and metastasis in ESCC.

Objective: Early prostate cancer micrometastatic foci undergo a mesenchymal to epithelial reverting transition, not only aiding seeding and colonization, but also rendering the tumor cells generally chemoresistant. We previously found that upregulated E-cadherin in the epithelial micrometastases activated canonical survival pathways, including PI3K-Akt, that protected the tumor cells from death; however, the extent of protection from blocking the pathway in its entirety was modest, because different isoforms may have alternately affected cell functioning. Here, we characterized Akt isoform expressions in primary and metastatic prostate cancers, as well as their individual contributions to chemoresistance. Methods: Akt isoforms and E-cadherin were manipulated with drugs, knocked down, and over expressed. Tumor cell killing was determined in vitro and in vivo. Overall survival was calculated from patient records and specimens. Results: Pan-Akt inhibition sensitized tumor cells to chemotherapy, and specific blockade of Akt1 or/and Akt2 caused cells to be more chemoresponsive. Overexpression of Akt3 induced apoptosis. A low dose of Akt1 or Akt2 inhibitor enabled standard chemotherapies to significantly eradicate metastatic prostate tumors in a mouse model, acting as chemosensitizers. In human specimens, we found Akt1 and Akt2 positively correlated, whereas Akt3 inversely correlated, with the overall survival of prostate cancer patients. Akt1high/Akt2high/Akt3low tumors had the worst outcomes. Conclusions: E-cadherin-induced activation of Akt1/2 isoforms was the essential mechanism of chemoresistance, whereas Akt3 made cells more fragile. These findings emphasized the need to target Akt1/2, rather than pan-Akt, as a rational therapeutic approach.

Background To investigate the effectiveness of tibiotalocalcaneal arthrodesis with a retrograde nail and allograft in limb salvage surgery for patients with distal tibia osteosarcoma. Methods 5 patients diagnosed as distal tibia osteosarcoma underwent ankle arthrodesis with a retrograde nail in our hospital. During the follow-up, radiographic views of the ankle joint were taken in two planes to assess bone healing and axis alignment. Other measurements of outcomes included procedure-related complications, local recurrence, and metastasis. Functional outcomes were evaluated with the Musculoskeletal Tumor Society (MSTS) scoring system. Results Postoperative complications occurred in 4 patients, including 4 cases of mild subcutaneous fluid and 1 case of screw breakage. All patients showed stable ankle and could stand or walk with the assistance of crutch before the complete union between allograft and host bone. One patient died due to multiple bone and pulmonary metastasis at 1 year after surgery. As for the other 4 patients, they were followed-up regularly for a mean period of 42 months. No local recurrence or distant metastasis occurred in any of these four patients. All the 4 patients expressed satisfaction with the outcome. According to MSTS scale, the mean postoperative functional score was 74.3%±4.4% (range, 70%–81%). Conclusions Intramedullary retrograde nail for distal tibia osteosarcoma could produce a satisfactory outcome in terms of functional results and complications. Excellent stabilization of the ankle joint can be achieved through this technique, as it allows patients to perform much earlier postoperative weight-bearing exercise.

While atypical expression of special AT-rich sequence-binding protein 2 (SATB2) has been approved associated with tumor progression, metastasis and unfavourable prognosis in various carcinomas. However, in oral squamous cell carcinoma (OSCC), both the expressive state and associated functions of SATB2's are still undefined. Here we show that, in clinical samples from a retrospective cohort of 58 OSCC patients, high expression of SATB2 is associated with poor prognosis of OSCC patients. In this study, we investigated SATB2 is highly expressed in OSCC tissues and cell lines, which can promote OSCC cells' proliferation, migration, invasion and tumor growth. According to sequencing results based on previous literature, we identified NOX4 is a bona fide downstream target of SATB2, when it was knockdown, OSCC's proliferation can be partially suppressed. Furthermore, NOX4 knockdown inhibits tumorigenicity, which can be rescued partially by ectopic expression of SATB2 in HNSCC cell line, and vice versa. Collectively, our findings not only indicate overexpression of SATB2 triggers the proliferative, migratory and invasive mechanisms which are important in the malignant phenotype of OSCC, but also identify NOX4 as the downstream gene for SATB2. These findings indicate that SATB2 may play a key role in OSCC tumorigenicity and may be a future target for the development of new therapeutic regimens.

Background Epidermal growth factor-containing fibulin-like extracellular matrix protein 2 (EFEMP2), also known as fibulin-4, MBP1 and UPH1, is an extracellular matrix protein associated with a variety of tumors. The purpose of this study was to investigate the prognostic value and the function of EFEMP2 in lung cancer. Methods The mRNA and protein expression of EFEMP2 in lung normal and cancer tissues, lung cancer cell lines (A549, H460, H1299 and H1650) and normal epithelial cell line BEAS-2B were evaluated by immunohistochemistry, RT-qPCR and Western blotting. The Public databases (Oncomine and Kaplan-Meier plotter) were used to investigate the prognostic value of EFEMP2 in lung cancer. RNA interference (RNAi) and overexpression transfection were performed to detect the effects of EFEMP2 up- or down-regulation on lung normal and cancer cell proliferation, invasion and metastasis in vitro and in vivo. Results EFEMP2 was lowly expressed in lung cancer tissues and cells, and its low expression was associated with malignant phenotype and poor prognosis of lung cancer. The same conclusion had been drawn from the Public databases. EFEMP2 overexpression significantly inhibited the invasion of lung cancer cells, hampered the process of EMT, and decreased the expression and activity of MMP2 and MMP9, while EFEMP2 knockdown remarkably enhanced the invasion of lung cancer cells, promoted EMT, and increased the expression and activity of MMP2 and MMP9. Conclusion The low expression of EFEMP2 was detected in lung cancer and was positively correlated with the poor prognosis of patients. EFEMP2 was a tumor suppressor gene that inhibited the progress of lung cancer, which suggested a new research objective for the future studies.

Mucin1 (MUC1) expression correlates with invasion and metastasis and poor survival in some cancers. The purpose of the study was to investigate the clinical significance of MUC1 expression and the risk of tumor metastatic recurrence in patients with esophageal squamous cell cancer (ESCC) after curative resection. A total of 108 ESCC patients were enrolled in this study. MUC1 expression was detected in ESCC tissues from 70 patients by immunohistochemistry (IHC). The expression of MUC1 in the cancerous tissue group was significantly higher than that in the paracancerous normal tissue group (65.4%:10.0%, p < 0.01). MUC1 expression correlated with pT (< 0.05), pN (p < 0.01) and pTNM stage (< 0.01). The 5-year survival rate of the patients was 39.8%. The 5-year tumor metastatic recurrence rate of the patients was 74.1%, and it was associated with pT (p < 0.01), pN (p < 0.01), pTNM stage (p < 0.01) and MUC1 expression (p < 0.01). Multivariate analysis confirmed that pN and MUC1 expression were independent predictive factors. In conclusion, MUC1 expression correlates with tumor metastatic recurrence in postoperative ESCC patients.

To develop and validate a clinical-radiomics nomogram for preoperative prediction of lung metastasis for colorectal cancer (CRC) patients with indeterminate pulmonary nodules (IPN). 194 CRC patients with lung nodules were enrolled in this study (136 in the training cohort and 58 in the validation cohort). To evaluate the probability of lung metastasis, we developed three models, the clinical model with significant clinical risk factors, the radiomics model with radiomics features constructed by the least absolute shrinkage and selection operator algorithm, and the clinical-radiomics model with significant variables selected by the stepwise logistic regression. The Akaike information criterion (AIC) was used to compare the relative strength of different models, and the area under the curve (AUC) was used to quantify the predictive accuracy. The nomogram was developed based on the most appropriate model. Decision-curve analysis was applied to assess the clinical usefulness. The clinical-radiomics model (AIC = 98.893) with the lowest AIC value compared with that of the clinical-only model (AIC = 138.502) or the radiomics-only model (AIC = 116.146) was identified as the best model. The clinical-radiomics nomogram was also successfully developed with favourable discrimination in both training cohort (AUC = 0.929, 95% CI: 0.885–0.974) and validation cohort (AUC = 0.922, 95% CI: 0.857–0.986), and good calibration. Decision-curve analysis confirmed the clinical utility of the clinical-radiomics nomogram. In CRC patients with IPNs, the clinical-radiomics nomogram created by the radiomics signature and clinical risk factors exhibited favourable discriminatory ability and accuracy for a metastasis prediction. • Clinical features can predict lung metastasis of colorectal cancer patients. • Radiomics analysis outperformed clinical features in assessing the risk of pulmonary metastasis. • A clinical-radiomics nomogram can help clinicians predict lung metastasis in colorectal cancer patients.

The poor prognosis of esophageal squamous cell carcinoma (ESCC) is mainly attributed to higher lymphatic-related metastatic ability. Whether the loss of expression of the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is associated with lymphatic-related metastasis needs elucidation. In the present study, we assessed the mRNA and protein level of PTEN in ESCC by qRT-PCR and immunohistochemistry. The results showed PTEN mRNA level in tumors was significantly lower than that in corresponding non-tumor esophageal epitheliums (p

Brain-specific metastasis is one of the primary causes of recurrence following complete resection of non-small cell lung cancer (NSCLC) and the underlying mechanism remains unclear. The present study was designed to investigate the correlation between C-X-C chemokine receptor type 4 (CXCR4) expression and brain-specific metastasis of NSCLC. Lung cancer tissues from 105 patients who underwent complete tumor resection between January 1998 and June 2008 (sample group, 34 with brain metastasis during the follow-up period; control group 1, 34 without metastasis during the follow-up period; and control group 2, 37 with other organ metastasis, excluding brain metastasis, during the follow-up period) were examined by immunohistochemistry to detect the expression of CXCR4 protein. The differences in CXCR4 expression were compared using McNemar’s χ2 test. Estimation of survival was calculated with the Kaplan-Meier method and the statistical differences were analyzed with the log-rank test. Overexpression of CXCR4 protein was observed in 31 (91.2%) NSCLC patients with brain metastasis, which was greater than that observed in the NSCLC patients with other organ metastases (73.0%; P=0.048) and without metastases (14.7%; P

Lung cancer is the leading cause of cancer‑related mortality worldwide. Over half of lung cancer cases are diagnosed after metastasis, for which the median survival time is approximately 8 months. microRNAs (miRNAs), which are a class of single‑stranded endogenous non‑coding RNAs, are likely to be involved in most biological processes. miR‑133 plays roles in cardiac development and disease, and recent studies showed that miR‑133 is downregulated in various human malignancies, such as bladder and lung cancer. However, its detailed role in the processes of cancer remains to be determined. In the present study, we found that in the lung cancer cell lines A549 and NCI‑H1299 overexpression of miR‑133a suppressed cell proliferation, migration and invasion. The miR‑133a‑induced suppression of cell migration and invasion can be reversed by miR‑133a‑specific inhibitor. According to the mRNA sequence, matrix metalloproteinase (MMP)‑14, which is an important regulator of metastasis, is a predicted target of miR‑133a. This was confirmed by dual luciferase reporter assay. Moreover, miR‑133a overexpression decreases the mRNA and protein levels of MMP‑14. Collectively, these results suggest that miR‑133a may inhibit lung cancer metastasis by targeting MMP‑14 and may be used as an anti‑metastatic therapy in lung cancer patients.

BACKGROUND To investigate the overexpression of Metastasis-associated gene 1(MTA1) protein and its relationship to the prognosis in esophageal squamous cell cancer after esophagectomy. METHODS 174 patients with middle third squamous cell carcinoma of the esophagus underwent complete resection in Provincial Hospital Affiliated to Shandong University between January 2002 and January 2005. The overexpression of MTA1 protein was detected by immunohistochemistry. Kaplan-Meier method was performed to calculate the survival rate, Cox regression multivariate analysis was performed to determine independent prognostic factors. RESULTS MTA1 protein overexpression rate in T1, T2, and T3 patients was separately 25.0, 31.9, and 53%, the difference of MTA1 protein overexpression between them was statistically significant (p = 0.017). The overexpression of MTA1 protein in patients with lymph node metastasis was significantly higher than those without metastasis (p = 0.042). MTA1 protein overexpression correlated with significantly worsened 5-year survival for all patients as well as those with T2 and T3 tumors, N0 nodal status or N1 nodal status. However, no significant correlations with T1 patients (p = 0.061). The result of Cox analysis demonstrated that N stage and MTA1 protein overexpression were independent prognostic factors. CONCLUSION MTA1 protein overexpression was detected in esophageal squamous cell carcinoma and was found to be significantly associated with the T stage. The patients with MTA1 protein overexpression had a significantly lower 5-year survival rate than without MTA1 protein overexpression. Lymph node metastasis and MTA1 protein overexpression were independent prognostic factors.

Introduction: Lymph node status is one of the most important prognostic factors for oesophageal squamous cell carcinoma (ESCC). Recent studies have revealed that several genes and molecules are involved in the origin and progression of ESCC. Tumour progression and metastasis require angiogenesis. It has recently been reported that vascular endothelial growth factor C (VEGF-C) induces not only angiogenesis but also lymphangiogenesis via VEGF receptor-2 and VEGF receptor-3. Aim: To investigate the correlation between VEGF-C expression and tumour metastatic recurrence in patients with ESCC. Material and methods: Ninety-two ESCC specimens were reevaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) to detect VEGF-C mRNA. The correlation between VEGF-C expression and clinicopathological factors was analysed using Fisher’s exact probability test or χ2 test. The logrank test was used to calculate the recurrence difference. And Cox regression multivariate analysis was performed to determine independent prognostic factors. Results: The VEGF-C mRNA expression was correlated with pT, pN, pTNM and lymphatic invasion. The 5-year tumour metastatic recurrence rate in patients after operation was significantly associated with pT, pN, pTNM stage and VEGF-C mRNA expression. The 5-year tumour metastatic recurrence rate of patients with VEGF-C mRNA expression in oesophageal cancer tissues was significantly higher than that of the patients without VEGF-C mRNA expression. The results of Cox regression multivariate analysis confirmed that pT status and VEGFC mRNA expression were independent relevant factors. Conclusions: The VEGF-C expression is related to tumour invasion, lymphatic invasion, lymph node metastasis, tumour stage and tumour metastatic recurrence in ESCC patients.

Brain-specific metastasis occurs frequently in lung cancer, and the mechanism is still unclear. The present study was designed to investigate the correlation between CXCR4 expression and brain-specific metastasis of non-small cell lung cancer. The brain metastatic tumors and lung cancer tissues from 32 patients with solitary brain metastasis of non-small cell lung cancer (M1 group), who underwent combined surgical treatment from January 1998 to June 2008, and 32 paired patients without distant metastasis (M0 group) and 30 patients with primary brain tumor, were examined by immunohistochemistry to detect the expression of CXCR4 protein. The difference of CXCR4 expression was compared by the McNemar χ2 test or Fisher’s exact test. Estimation of survival was calculated with the Kaplan–Meier method, and the statistical differences were analyzed with the log-rank test. Overexpression of CXCR4 protein was observed in 29 (90.6%) M1 non-small cell lung cancers and in all (100%) brain metastatic tumors, which was significantly higher than that in the paired M0 non-small cell lung cancer and the primary brain tumors, respectively (p = 0.000). The 3- and 5-year cumulative survival rates of patients with solitary brain metastasis of lung cancer were 21.9 and 12.5%, significantly lower than the corresponding survival rates of M0 group patients (p = 0.005). CXCR4 protein was highly overexpressed in M1 non-small cell lung cancer and brain metastatic tumors, which indicated that high-level CXCR4 expression correlates with brain-specific metastasis of non-small cell lung cancer.

Calreticulin is an essential, multifunctional Ca(2+)-binding protein that participates in the regulation of intracellular Ca(2+) homeostasis, cell adhesion, and chaperoning. Calreticulin is abundantly expressed and regulated by androgens in prostate epithelial cells. Given the importance of both calreticulin in multiple essential cellular activities and androgens in prostate cancer, we investigated the possibility of a role for calreticulin in prostate cancer progression. Immunohistochemistry revealed the down-regulation of calreticulin in a subset of human prostate cancer specimens. Prostate cancer cells overexpressing exogenous calreticulin produced fewer colonies in both monolayer culture and soft agar. Furthermore, calreticulin overexpression also inhibited tumor growth in the orthotopic PC3 xenograft tumor model and macroscopic lung metastasis in the rat Dunning AT3.1 prostate tumor model. To address the potential mechanism of calreticulin suppression of prostate cancer, we generated calreticulin mutants with different functional domains deleted. The calreticulin mutants containing the P-domain, which binds to other endoplasmic reticulum chaperone proteins, were sufficient for the suppression of PC3 growth in colony formation assays. Overall, our data support the hypothesis that calreticulin inhibits growth and/or metastasis of prostate cancer cells and that this suppression requires the P-domain.

The high recurrence rate after surgery is the main reason for the poor prognosis of esophageal squamous cell carcinoma (ESCC) patients. Finding indicators of recurrence and taking adjuvant therapy may be useful for patients in high risk of recurrence. claudin-4 (CLDN4) is the core protein to form the tight junction, which plays an important role in cell adhesion, and its aberrant expression were detected in various cancers while its expression and functions in ESCC still remained unclear. Here, we detected the expression of CLDN4 in 114 ESCC tissue samples by real-time RT-PCR and immunohistochemistry, and the result showed that the low expression of CLDN4 correlated with a higher T staging (P = 0.010), lymphatic metastasis (P < 0.001) and recurrence status (P = 0.002). And the Cox regression analysis showed that the T classification (P = 0.005), lymph node metastasis (P = 0.003) and low CLDN4 expression (P = 0.029) were independent risk factors of recurrence. Further, we proved the CLDN4 in inhibiting growth, colony formation and invasion in vitro by establishing two stable CLDN4-silencing ESCC cell lines. In conclusion, CLDN4 played an important role in preventing metastasis and could be an effective biomarker to predict the recurrence of ESCC.

Objective To investigate the association between v-Ki-ras2 Kirsten rat sarcoma viral oncogene homologue ( KRAS) gene mutations and levels of human leucocyte antigen (HLA) class I antigen in primary lung tumours and metastatic lymph nodes of patients with non-small cell lung cancer (NSCLC). Methods Patients with NSCLC undergoing tumour resection were enrolled. KRAS codon 12 mutations were analysed in normal lung and lymph node tissue, primary lung tumours and metastatic lymph nodes using polymerase chain reaction–restriction fragment length polymorphism analysis. HLA class I antigen immunostaining was examined using flow cytometry. Results A total of 65 patients participated in the study. All normal lung tissues had positive HLA class I antigen immunostaining. The majority of primary lung tumours (56/65) and all of the metastatic lymph nodes (31/31) had downregulated HLA class I antigen immunostaining. There was a positive correlation between downregulated HLA class I antigen in primary tumours and metastatic lymph nodes. There was a negative correlation between KRAS codon 12 mutations and the level of HLA class I antigen in primary and metastatic tumours. Conclusions KRAS codon 12 mutations appear to be important in the downregulation of HLA class I antigen in NSCLC. Abnormal activation of the oncogenic KRAS pathway might provide a new treatment target for NSCLC.

The purpose of the present study was to investigate the risk factors associated with lymph node metastatic recurrence in patients with N0 esophageal cancer after Ivor-Lewis esophagectomy based on the detection of Mucin 1 mRNA and vascular endothelial growth factor (VEGF) C mRNA. The subjects were 82 patients with pN0 esophageal cancer who underwent Ivor-Lewis esophagectomy with two-field lymph node dissection from January 2001 to January 2005. A total of 501 lymph nodes obtained from these patients were re-evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) to detect mucin l (MUC1) mRNA; VEGF-C mRNA was also detected in esophageal cancer issues by RT-PCR .The diagnosis of lymph node micrometastasis (LNMM) was based on the detection of MUC1 mRNA. The Kaplan–Meier method was used to calculate the survival rate and lymph nodal metastatic rate, the log-rank test was performed to compare the recurrence rate, and Cox regression multivariate analysis was performed to determine independent prognostic factors. MUC1 mRNA was detected in 29 lymph nodes from 23 patients, which accounted for 5.79% of all the 501 lymph nodes and 28.05% of all 82 patients, respectively. Vascular endothelial growth factor C mRNA was identified in esophageal cancer issues from 42 (51.22%) patients. The overall 3- and 5-year survival rates of 82 patients were 78.0 and 51.2%, respectively. First recurrence exhibiting lymph nodal metastasis was recognized in 37 patients (45.1%) in the first 3 years after operation. The lymph node metastatic rate in patients in the first 3 years after operation was significantly associated with T status (p

Metastasis-associated protein 1 (MTA1 protein) has been reported to be correlated with the biological behavior and prognosis of several malignant carcinomas. We hypothesized that stage I non-small cell lung cancer (NSCLC) patients with MTA1 protein overexpression would be more likely to have a poor prognosis. Therefore, we tested the expression of MTA1 protein in 60 stage I NSCLC and 30 paracarcinous normal lung tissues using the streptavidin-perosidase method. The Kaplan-Meier method was used to calculate the survival rate, and Cox regression analysis was performed to identify prognostic risk factors. MTA1 protein overexpression was detected in 22 stage I NSCLC tissues in this study. Tumor differentiation and tumor diameter were significantly associated with MTA1 protein overexpression, while not correlated with age, sex, pathological type or smoking status. The five-year survival rate of patients with MTA1 protein overexpression was significantly lower than that of those without expression (40.9% vs. 84.1%; P

The purpose of the present study was to investigate the expression of metastasis-associated protein 1 (MTA1) and its relationship to the disease-free interval after resection of pathologic N0 (pN0) esophageal squamous cell cancer (ESCC). The subjects were 90 patients who successfully underwent complete resection of pN0 ESCC between May 2001 and May 2003. Immunohistochemical staining for MTA1 protein was performed using the avidin–biotin peroxidase complex method. Log-rank test was performed to compare the disease-free interval, and Cox regression multivariate analysis was performed to judge independent prognostic factors. Metastasis-associated protein 1 overexpression was detected in 40 esophageal cancer tissues. Disease-free interval was significantly associated with MTA1 protein overexpression (p = 0.015). The overall 5-year survival rate was 45.6%, the 5-year survival rate of patients with MTA1 protein overexpression was significantly lower than that of those without overexpression (25.0 versus 62.0%; p

Objective: Mediastinal lymph node metastasis (N2) is a key prognostic factor for lung carcinoma. This study was undertaken to investigate the relationshipbetweenvascularendothelial growthfactorC (VEGF-C)expressionandpostoperativeearly recurrence in patientswith N2 non-smallcell lung cancer. Methods: Cancer tissue samples from 92 patients with pN2 non-small-cell lung cancer and benign lung disease tissues samples from 30 patients were examined by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry assays to detect VEGF-C expression. The difference of VEGF-C expression was compared by x 2 test. All patients with N2 disease were evaluated within 1 year after surgery to detect early tumour recurrence. Cox regression analysis was performed to determine the risk factors of postoperative early recurrence of N2 lung cancer. Results: VEGF-C mRNA expression was observed in 64 (70%) pN2 lung cancer tissues, but was not found in benign lung disease tissues. Early recurrence occurred in 43 patients (47%) at 1 year after operation. The main pattern was distant recurrence, and the most frequent sites were the brain and lung. The early recurrence rate in patients with positive VEGF-C expression was significantly higher than that of those with negative VEGF-C expression (P = 0.006, log-rank test). Cox regression analysis revealed that positive VEGF-C expression in tumours (hazard ratio (HR) = 2.523, P = 0.037) was an independent risk factor of postoperative early recurrence of N2 lung cancer. Conclusions: VEGF-C expression was high in N2 lung cancer, with significant correlation to postoperative early recurrence. About one-half of the patients with N2 non-small-cell lung cancer would develop recurrence disease within 1 year after surgery, frequently with mediastinal nodes, brain or lung metastases. VEGF-C might be a predictor of postoperative early recurrence in patients with N2 non-small-cell lung cancer. # 2009 European Association for Cardio-Thoracic Surgery. Published by Elsevier B.V. All rights reserved.

Androgens are intimately associated with prostate cancer progression. We have previously identified more than 24 androgen-response genes. One of the genes encodes calreticulin, a highly conserved protein with demonstrated functions in intracellular Ca(++) homeostasis, cell adhesion, chaperoning, and gene expression. Our studies have indicated that calreticulin overexpression is suppressive to anchorage-independent growth and metastasis of prostate cancer cells and calreticulin expression is down-regulated in human prostate tumor specimens. Thus, down-regulation of calreticulin in clinical prostate cancer specimens is likely to be an important step in prostate cancer progression. Our observations argue that part of androgen-induced gene expression program, such as calreticulin, is inactivated in the progression of prostate cancer, which represents a new concept in prostate cancer biology. Our results also provided strong basis for further exploring the mechanism by which calreticulin suppresses prostate tumor metastasis. In addition, we have generated several mutants for calreticulin, which will allow us to determine which of the three domains, N, P, or C, is responsible for the suppression of prostate tumor metastasis.