Relationship Of MiR-221 Expression With The Stage, Type And Prognosis Of Hepatocellular Carcinoma

Aim:Hepatocellular carcinoma（HCC） is a common malignant cancer in our country and screening related genes functioned in HCC genesis and development may give some basis to diagnosis and treatment of this disease. micro RNA（mi RNA） is a kind of intrinsic, 18-22 nt long, non-coding RNA. It functions its biological roles by inhibiting expression of target genes on post-transcriptional level. Studies showed that mi RNA may correlates with HCC genesis and development. The present study tested the expression of mi R-221 in HCC tissues using q PCR, and then statistic the relationship of mi R-221 expression with the stage, type and prognosis of Hepatocellular carcinoma. We further investigated the change on biological characters of HCC cells after blocking the expression of mi R-221.Methods:After collected 246 cases of HCC and paired paratumor tissues, we tested the expression of mi R-221 using q PCR and then grouped all the cases into two groups according to the expression of mi R-221 as high expression group and low expression group, respectively. Next, we analized the relationship of mi R-221 expression and HCC clinical parameters using survival statistical methods, such as kaplamier, Cox proportional hazard regression model etc. In our in vitro study, we designed the antisense sequence of mi R-221, and transfected HCC cell line by lentivirus vectors. Then the proliferation, clone, invasion and metastasis of the cells were examined by CCK-8, cloning test, wound healing and transwell assay, as well as flowcytometry. Finally, western blots were also used to test the cell cycle proteins, p27^kip1 and p57^kip2 after mi R-221 blocking.Results:Expression of mi R-221 is correlated with pathological grades（P=0.0001）, TNM stages（P=0.03）, portal vein tumor thrombus（P=0.001）, AFP（P=0.007） and lymphocyte metastasis（P=0.001）. But mi R-221 had no relationship with age, sex, tumor size, tumor count and diolame formation. mi R-221 high expression is negatively related with HCC overall survival. Results of Cox proportional hazard regression model showed that mi R-221 is an independent factor for HCC prognosis. Blocking mi R-221 expression significantly inhibited the proliferation, cloning, invasion and metastasis of HCC cell line. The cell cycle arrest of HCC cells on G1 stage was induced by mi R-221 expression blockage. Western blot data showed that p27^kip1 and p57^kip2 were significantly increased after blocking mi R-221 expression.Conclusion: High expression of mi R-221 may be associated with HCC tumor genesis and development, and be an independent negative prognostic factor in HCC. Blockage on of mi R-221 in HCC cells may shade a novel light on the target for diagnosis and treatment of HCC.