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Wei Huang, Feizhou Huang, Zhao Lei, Hongwu Luo Department of Hepatopancreatobiliary Surgery, The Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, People’s Republic of ChinaCorrespondence: Hongwu LuoDepartment of Hepatopancreatobiliary Surgery, The Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, People’s Republic of ChinaTel/Fax +86 731 8861 8576Email lhwxy123456@163.comBackground: The most common malignant tumor of the digestive system is HCC. However, the mechanism and pathogenesis of HCC occurrence and progress are still unknown. LncRNA is closely related to the occurrence and progress of HCC. It is important to investigate the effect and role of lncRNA in HCC.Materials and Methods: LncRNA microarray assay was used to screen the differential expression profile of lncRNA. SNHG11, miR-184 and GO2 expression was analyzed by RT-PCR. The ability of SNHG11 to serve as a sponge for miRNA and the fact that miR-184 directly targets mRNA were revealed by dual luciferase assay and RIP. Apoptosis and autophagy related proteins were detected by Western blot. Cell proliferation, invasion, migration, and apoptosis were detected by CCK-8 assay, wound healing assay, transwell assay, and flow cytometry.Results: LncRNA microarray assay and RT-PCR results revealed that the expression of SNHG11 was increased in HCC tumor tissues and also upregulated in HCC cells. SNHG11 had a connection with poor survival rate in HCC. In addition, dual luciferase assay and RIP results revealed that SNHG11 serves as a sponge for miR-184 and miR-184 directly targets AGO2. Pearson correlation analysis showed that SNHG11 with miR-184 and miR-184 with AGO2 were negative correlations, and SNHG11 with AGO2 was a positive correlation. Cell function assay and Western blot showed SNHG4/miR-184/AGO2 regulatory loop was critical for HCC cell proliferation, migration, apoptosis, and autophagy.Conclusion: Our study demonstrated that the expression of SNHG11 is higher in HCC; moreover, SNHG11 promotes proliferation, migration, apoptosis, and autophagy by regulating AGO2 via miR-184 in HCC. Our verification of the role of SNHG11 may provide a novel biomarker for the diagnosis, therapy, and prognosis of HCC.Keywords: hepatocellular cancer, LncRNA SNHG11, apoptosis, biomarker

Lei Zhu,1 Feizhou Huang,1 Tao Wan,1 Hongbo Xu,1 Qian Zhao2,3 1Department of Hepatobiliary and Pancreatic Surgery, the Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, People’s Republic of China; 2Department of Adult Stem Cells, Institute of Reproduction and Stem Cell Engineering, Central South University, Changsha, Hunan 410078, People’s Republic of China; 3Institute of Transformation Medicine Affiliated to Hunan Yearth Biotechnology Co., Ltd., Changsha, Hunan 410200, People’s Republic of China Introduction: Hepatocellular carcinoma (HCC) is the third major cause of malignant tumor-related death worldwide because it is initially diagnosed in the advanced stage and its therapeutic outcomes are usually poor. Based on this, it is urgent to identify effective early diagnosis biomarkers and new therapeutic targets to promote HCC treatment. Long noncoding RNAs (lncRNAs) have been reported as promising biomarkers for tumor diagnosis and treatment.Materials and methods: In this study, we profiled expression patterns and dysregulation of lncRNAs in HCC tissues by analyzing two datasets GSE55191 and GSE64631 from Gene Expression Omnibus database firstly, each of which contains expression profiles of 3 primary HCC tissues and 3 normal liver tissues, respectively.Results: LncRNA 882 (LINC00882) is one of the lncRNAs that was significantly upregulated in HCC tissues compared with normal liver tissues. We verified the upregulation of LINC00882 in HCC by using two separate cohorts that contained 85 HCC tissues and paired adjacent noncancerous tissues and 86 HCC tissues and 89 independent noncancerous liver tissues, respectively.Discussion: We found that upregulation of LINC00882 is correlated with poorer prognosis of HCC patients. In vitro cell experiments demonstrated that knockdown of LINC00882 inhibits proliferation, migration and invasion of HCC cell lines.Conclusion: These results indicated that LINC00882 promotes HCC progression and could be a potential prognostic biomarker and therapeutic target for HCC. Keywords: hepatocellular carcinoma, long noncoding RNAs, LINC00882, proliferation, migration

The incidence and mortality of liver cancer increased year by year. Our country presents high incidence of liver cancer. MicroRNAs have tissue sensitivity as tumor biomarkers that play a role by promoting tumor growth as oncogenes or inhibit malignant cell growth as tumor suppressor genes. Studies showed that miR-15b abnormal expression in the tumor and can be treated as one of the tumor molecular markers. However, miR-15b expression and role in the liver cancer cells have not been elucidated. This study intended to explore the mechanism of miR-15b effect on liver cancer occurrence and development. Liver cancer cell line HepG2 was transfected with miR-15b mimic or inhibitor. Real-time PCR was applied to detect miR-15b expression. MTT was used to test cell proliferation. Transwell assay was performed to determine cell invasive ability. Real-time PCR and Western blot were used to detect BCL2 expression. MiR-15b mimic transfection promoted miR-15b overexpression and inhibited HepG2 cell proliferation significantly (P < 0.05). MiR-15b overexpression downregulated BCL2 mRNA and protein expression obviously (P < 0.05). On the contrary, miR-15b inhibitor transfection markedly reduced miR-15b expression in liver cancer cells (P < 0.05), promoted tumor cell proliferation, and increased BCL2 mRNA and protein expression. MiR-15b expression changes did not affect cell invasion (P > 0.05). MiR-15b can inhibit HepG2 cell proliferation and down-regulate BCL2 mRNA and protein expression.

TRAIL (TNF-related apoptosis-inducing ligand) is a member of the tumor necrosis factor superfamily that can induce tumor selective death by up-regulating death receptor 4 (DR4) and DR5 expression. The study aimed to explore the role of RIP and c-FLIP genes in TRAIL induced liver cancer cell HepG2 and Hep3B apoptosis and related mechanism. RIP and c-FLIP silenced HepG2 and Hep3B cell model were established through siRNA. Western blot was applied to test c-FLIP, RIP, DR4, DR5, FADD, Caspase-3/8/9, ERK1/2, and DFF45 protein expression. Caspase-8 kit was used to detect Caspase-8 expression. Flow cytometry was performed to measure cell apoptosis rate. Acid phosphatase method was applied to determine cell cycle. TRAIL had no significant effect on Caspase-3/8/9, DR4, DR5, ERK1/2, and DFF45 protein expression, but up-regulated c-FLIP and RIP protein expression and reduced FADD expression level. After treated by the chemotherapy drug mitomycin and adriamycin, c-FLIP and RIP expression decreased significantly, while FADD increased. After knockout c-FLIP and RIP gene, HepG2 and Hep3B cell apoptosis rate induced by TRAIL increased obviously. Meanwhile, cell subG1 percentage increased markedly and exhibited G1 phase growth retardation. In addition, after two kinds of gene knockout, Caspase-8 was activated and produce Caspase-3 P20 and P24, leading DFF45 appeared DNA fragment P17 and P25. c-FLIP and RIP can inhibit Caspase-8 activation and prompting HepG2 and Hep3B resistant to cell apoptosis induced by TRAIL.

Objective: This study aimed to define the correlation between microRNA-122 (miR-122) expression and hepatectomy-induced liver injury in patients with hepatocellular carcinoma (HCC). Methods: Plasma miR-122 expression in patients with HCC and healthy age-matched controls was determined, and correlated with plasma alanine transaminase activity (ALT) and bilirubin levels preoperatively and on days 1 and 7 postoperation. Correlations between plasma miR-122 and clinicopathological characteristics at 1 day postoperation were also determined. Results: This study included 80 patients with HCC and 80 controls. Baseline expression of miR-122 mRNA and ALT in patients with HCC was significantly higher than in controls. MiR-122 expression correlated with ALT and bilirubin levels preoperatively and on days 1 and 7 postoperatively. In patients with HCC who received a block of the first hepatic portal during surgery and those with excised tumour size >5 cm, plasma miR-122 expression was significantly increased on day 1 postoperatively, compared with expression levels in those who did not receive a block and those with smaller tumours. Conclusions: Plasma miR-122 expression is correlated with hepatectomy-induced liver injury in patients with HCC. Increase in miR-122 expression could be used as an index of such injury before and after hepatectomy in these patients.

To study the role of perforating veins in predicting the likelihood of esophageal variceal rupture and variceal recurrences.In 70 patients with esophageal varices, a 20 MHz ultrasonographic transducer was used to image esophageal varices; the radius and perforating veins were calculated. Esophageal variceal pressure measurements were obtained by noninvasive pressure gauge. The relationship between the size of esophageal varices and the presence of perforating veins in the esophageal wall was studied using chi-square test; the patients were divided into two groups according to the presence of perforating veins, and the pressure in each group was compared by using Student's t test. In addition, the frequency of endoscopy sessions necessary for varix eradication, the dots of endoscopic variceal ligation and recurrence of esophageal varices within a year were also compared by the Mann-Whitney U test.The presence of perforating veins in the esophageal wall was significantly higher in patients with large radius of varices than in patients with small radius. The esophageal variceal pressure in patients with perforating veins was greater than that of patients without perforating veins (23+/- 4.5 vs 12+/-3.1 mmHg, p0.05). The frequency of endoscopy sessions required for varix eradication and the dots of EVL in patients with perforating veins was greater than that in patients without perforating veins (3.25 +/- 0.50 vs. 2.11+/-.78; 25 +/-.50 vs. 18.56+/- 5.46 p0.05). The recurrence of esophageal varices within a year was higher in patients with than in patients without perforating veins (75.93 vs. 18.75%, p0.05).Perforating veins in the esophageal wall correlate with the recurrence of esophageal varices in patients with portal hypertension