| Objective: HCC is one of the most common malignant tumors in China,which featues have high degree of malignancy, easy recurrence andmetastasis, and poor prognosis. The radiotherapy and chemotherapy againsthepatocellular carcinoma is not very effective, even if the Hepatectomy isimplemented in hepatocellular carcinoma patients,whose3-year survival rateis only40-50%. The reason is associated with biological characteristics ofinvasion and metastasis of liver cancer. Before,people thought that the featureof growth, invasion and metastasis of tumor is the result of the role of alltumor cells. But now, the study found that the invasion and metastasis oftumor were due to that of the stem cell-like cells that are parts of the tumorcells. These cells also has the characteristics of tumor cells and stem cells,that are tumor initiating cells, called cancer stem cells. So the biologicalcharacteristics of invasion and metastasis of cancer are closely related tocancer stem cells. tumor stem cells had been Isolated and identified fromleukemia, breast cancer, brain tumors and lung cancer. Suetsugu A, et al.isolated and identified CD133-positive cells from the liver cancer Huh7cells,that had stem cell characteristics with a high degree of tumorigenicity andproliferatiion potential to differentiattion, and that called liver cancer stemcells. Because out of control of the regulation of cell self-renewal capacity is akey factor for tumor development, studying genes that was related to themechanism of regulation of cell self-renewal in tumor cells was particularlyimportant. the stem cell-related gene OCT4is One of these genes. The studyfound abnormal expression of stem cell gene-related OCT4in hepatocellularcarcinoma. Had reported that OCT4was a potential cancer genes and played arole in the regulation of liver cancer stem cells, but few reports. Some studieshave speculated that the the regulation of cancer stem cells by OCT4may be achieved through the regulation of the Wnt signaling pathway. The subjectstudied the relationship between stem cell related gene OCT4and Wntpathway in HepG2cells by interfering with gene OCT4and the affect ofexpression of OCT4on liver cancer cell migration by scratch test study.Explore regulatory mechanism of the OCT4in the development of liver cancerand its affect on hepatocellular carcinoma cell biological characteristics,andprovide a theoretical basis to seek the feasibility of OCT4as a treatment forliver cancer targets.Method: Build OCT4-siRNA and the targeted design of the negativecontrol OCT4-a ncRNA (no effect) and Liposome-mediated transfect HepG2cells whose OCT4is of high expression.This experiment is divided into twogroups,OCT4-si group, the experimental group and OCT4-nc group,negative control group. OCT4-siRNA was transfected into HepG2cells24hlater,Appraise silencing efficiency of the OCT4gene by RT-PCR method.Observed wnt3and bete-catenin mRNA expression in the wnt signalingpathway when the OCT4gene is silenced in human HepG2cells.And,by thecell migration assay, detect the changes in capacity of cell migration of theliver cancer before and after the OCT4gene is silenced.Result:1OCT4mRNA expression levels were0.204±0.305in experimentalgroup and0.408±0.314in negative control group, and the expression levelof OCT4mRNA was significantly lower than the negative control group (P<0.05).2wnt3mRNA expression levels were0.243±0.003in experimentalgroup and0.337±0.025in negative control group,and the expression level ofwnt3mRNA was significantly lower than the negative control group (P<0.05).3bete-catenin mRNA expression levels were0.629±0.041inexperimental group and0.876±0.039in negative control group,and theexpression level of bete-catenin mRNA was significantly lower than thenegative control group (P <0.05). 4Scratch test results show that after24h, the cell migration rate of thenegative control group is50.67±5.67%and that in experimental group is only29.67±3.79%,which difference was significantly (P<0.05);48h later, in thenegative control group,cell migration rate was71.42±7.02%, while in theexperimental group, cells migrate rate wasonly70.81±8.73%, Nosignificant difference in cell migration rate in the experimental group and thenegative control group,no statistical significance (P>0.05)Conclusion:1By the OCT4-siRNA interference in liver cancer cells,compared withthe negative control group, OCT4mRNA expression was significantlydecreased in the experimental group. That descripted siRNA successfullytransfected to silence the expression of OCT4in HepG2cells.2Before Silencing the OCT4gene, OCT4mRNA in liver cancer cellshighly expressed and Wnt3mRNA and β-catenin mRNA of high expressiontoo, indicating that there may be some relationship between the OCT4and theWnt signaling pathway. After silencing OCT4gene, OCT4mRNA expressiondecreased, and the relative expression levels of Wnt3mRNA and of β-cateninmRNA in the Wnt signaling pathway also fell. This further explained thatthere is a certain relationship between the stem cell genes OCT4and Wntsignaling pathways. This prompted OCT4may be on the occurrence of livercancer development work through the regulation of the Wnt signalingpathway.3The scratch test: compared with the negative control group,transfected24h, liver cancer cell migration rate of the experimental group wassignificantly lower than the negative control group. Note that the abnormalexpression of OCT4was related to invasion and metastasis of liver cancer.48h after transfection,the liver cancer cell migration rates of the experimentalgroup and the negative control group were non-discrimination,possiblybecause of siRNA degradation.4According to this research,note that OCT4was related to invasion andmetastasis of liver cancer, and play an important role in the occurrence of liver cancer development, and OCT4may be as a target in the treatment of livercancer. |
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