The Study On The Expression And Biological Characteristics Of ACTL6A Gene In Cholangiocarcinoma

Objective: Cholangiocarcinoma is one of the most aggressive malignancies and itsprognosis is poor due to its early aggressive characteristics and poor response to chemotherapy or radiation therapy.A growing body of evidence suggests that aberrant expression ACTL6 A is involved in the development and progression of cancer.However,their clinical value and biological role in cholangiocarcinoma remains unclear.Our preliminary study found that ACTL6 A was abnormally expressed in cholangiocarcinoma tissues.Therefore,we speculate that the abnormal expression of ACTL6 A might be related to the occurrence and development of cholangiocarcinoma.This study was designed to investigate the biological role and potential mechanism of ACTL6 A in the occurrence and development of cholangiocarcinoma.Methods: First of all,we detected the expression of ACTL6 A in cholangiocarcinoma by Western blot.Secondly,RNA interference(RNAi)was used to mediate and transfect small interfering RNA(siRNA)into cholangiocarcinoma QBC939 cells by liposome.The experiment was divided into the negative control group(siRNA-NC)and the silencing group(siRNA-ACTL6A).After siRNA transfection,silencing efficiency was evaluated by Real-Time PCR(RT-PCR)and Western blot.Cell proliferation was detected by CCK-8assay and colony formation assay;scratch assay was used to detect cell migration ability.Transwell migration and invasion assays were used to detect cell migration and invasion;Real-Time PCR was used to detect the expression of epithelial marker factor E-cadherin and mesenchymal marker factor Vimentin and N-cadherin in cells;Western blot analysis expression levels of epithelial marker protein E-cadherin and mesenchymal marker protein Vimentin and N-cadherin in cells.Statistical analysis was performed using GraphPad Prism 6.0 software.Results: The expression of ACTL6 A in cholangiocarcinoma was significantly higher than that in the normal tissues.After ACTL6 A siRNA was transfected into cholangiocarcinoma QBC939 cells,the mRNA and protein expression levels of ACTL6 A in the silencing group were significantly decreased(p<0.05).After silencing the ACTL6 A gene in cholangiocarcinoma cells,CCK-8 results showed that the proliferation rate of cells was significantly decreased(p<0.05);The results of colony formation assay showed that the number of cell colonies in the silencing group was significantly decreased(p<0.05);the results of the scratch test showed that the migration ability of the silencing group was significantly weakened(p<0.05);Transwell migration and invasion experiments showed that the number of cells which could pass through the chamber was significantly reduced in the silencing group(p<0.05).These results indicated that the silencing of the ACTL6 A gene significantly inhibited cell migration and invasion;The results of Real-Time PCRshowed that silencing ACTL6 A expression can up-regulated the expression of epithelial marker factor E-cadherin and down-regulated the expression of mesenchymal marker factor Vimentin and N-cadherin(p<0.05).Furtherly,the results of Western blot showed that silencing of ACTL6 A gene can also up-regulated the expression level of epithelial marker protein E-cadherin and down-regulated the expression levels of mesenchymal marker protein Vimentin and N-cadherin(p<0.05).Conclusion: Our results indicate that ACTL6 A is abnormally expressed in cholangiocarcinoma tissues.After silencing the ACTL6 A gene,the proliferation ?migration and invasion of cholangiocarcinoma QBC939 cells are significantly inhibited.The expression of epithelial marker E-cadherin was up-regulated,while the expression of mesenchymal marker Vimentin and N-cadherin was down-regulated,which may be related to the inhibition of EMT.Therefore,it is speculated that ACTL6 A may be a potential new target for the prevention and treatment of cholangiocarcinoma.