The Mechanism Of MicroRNA-21 Expression On The Invasion And Metastasis Of Cholangiocarcinoma Cells

?Backworld? Cholangiocarcinoma is a malignant tumor originating from bile duct epithelial cells,including intrahepatic and extrahepatic cholangiocarcinoma,accounting for about 3% of gastrointestinal cancer.In recent years,with the continuous improvement of diagnostic level,the global incidence rate is on the rise,especially in our country.Cholangiocarcinoma is characterized by occult onset,rapid development,poor prognosis and so on.Surgical resection is recognised as the preferred method of cure for cholangiocarcinoma,but radical resection(including liver transplantation)is only about 10%,the median survival time is less than 24 months.Therefore,the development of new screening and diagnostic methods to find effective treatment of cholangiocarcinoma target,to improve the diagnosis and survival rate of patients with cholangiocarcinoma,control the recurrence rate,improve the prognosis has a very important clinical value.Micro RNA is an endogenous non-coding RNA of 19-25 nucleotides in lengthcan,play a new target of tumor gene therapy by regulating the signal transduction pathways involved in the target gene,affecting the occurrence and development of tumor and playing the biological function similar to oncogene or tumor suppressor gene.However,fewer miRNAs have been identified,so finding and identifying new miRNAs are oncogenes or tumor suppressor genes,plays a crucial role in directing targeted therapy.Recent studies have shown thatmicro RNA-21(abbreviated miR-21)is highly expressed in cholangiocarcinoma cells and has a sensitivity and specificity of more than 95% in cholangiocarcinoma tissues.Suggesting that miR-21 may be a new marker for early diagnosis,therapy of cholangiocarcinoma.In clinical practice,alternative cholangiocarcinoma tissue is very difficult to obtain,and have great lesion.Because of the noninvasive nature of circulating miRNA detection,it is expected to be a rapid,sensitive tumor diagnosis,prognostic analysis of molecular markers.Our recent study showed that miR-21 was up-regulated in cholangiocarcinoma tissues,which was significantly higher than that in adjacent non-tumor tissues,and was closely related to the degree of differentiation,intrahepatic metastasis,survival and prognosis of cholangiocarcinoma.More important is the discovery of cholelithiasis and cholangiocarcinoma in patients with bile miR-21 there is different expression.?Objective? To defect the expression of miR-21 in cholangiocarcinoma cells have effect on cell invasion and metastasis ability,and further explore its underlying mechanism.?Methods? Construction of lentiviral vector,miR-21 gene infected with cholangiocarcinoma RBE cells were established overexpression group,overexpression group,inhibition group,inhibition control group and blank control group.RT-PCR analyzes was used to detect of 5 groups of stable cell lines.The invasion and metastasis effect of RBE cells were detected by Transwell invasion assay and wound-heading array.The expression of E-cadherin,N-cadherin and Vimentin-related genes and the expression of PTEN gene were detected by Western blot.?Results?The invasion and metastasis ability of the infected group were significantly higher than those of the negative control group and the blank control group.In contrast,the invasion and metastasis ability of the miRNA group were significantly lower than that of the negative and blank group.And EMT-related proteins,in which N-cadherin and Vimentin were increased in overexpression group and E-cadherin expression was decreased.The effect of inhibitory group was opposite to that of overexpression group.The expression of PTEN protein in theoverexpression group was inhibited,and the expression of PTEN protein was decreased in the negative group and the blank group.?Conclusion? Overexpression of miR-21 enhances the ability of invasion and metastasis of cholangiocarcinoma cells,and down-regulation of miR-21 can lead to the invasion and metastasis of cholangiocarcinoma.MiR-21 may play an important role in the invasion and metastasis of cholangiocarcinoma cells by acting on umor suppressor gene PTEN and EMT-related proteins(E-cadherin,N-cadherin,Vimentin).