Study On Tumorigenesis Of Ovarian Cancer Cell Line Skov - 3 By Silencing ATF5

Objective:Analysis after silencingATF5, the effect on ovarian cancer cells proliferation and a poptosis in vitro and in vivo in SKOV-3cells, to gain a preliminary discussion ab out the application value of ATF5silence in human.Methods:1、Alentiviral vector with RNA interference(RNAi) of ATF5gene infected SKOV-3cells.compared with an empty vector transfection SKOV-3cells and normal SKO V-3cells, in order to observe the growth of the three groups of cells proliferation.2、MTT method is used to detect culture cell proliferation in vitro growth.3、We used Situ Cell Death Detection Kit(TUNEL) to detect cell apoptosis.4、Plant3kinds of cells in nude mice respectively,observe each group survival c ondition、tumor growth in nude mice, and calculate tumor rate.5、RT-PCR is used to detecte the expression of ATF5mRNA in tumor tissue.6、Western Blot is used to detecte the expression of ATF5protein in tumor tissue.7、HEstaining is used to observe tumor biological change in the form of organizat ion.Results:1、There was no significant difference in vitro culture cell biology form during thr ee groups, at the same time the cell proliferation has no statistical significance.Th e group with ATF5-silence transfection has a higher apoptosis than the blank con trol group and the nonspecific transfection group.2、After inoculation, there was no significant difference in the three groups of nud e mouse tumor rate。Compared with the other control groups, the tumors of specif icity transfection group grew later.3、Tumor tissue rt-pcr showed there was statistically significant difference in ATF5mRNA expression (P<0.05).4、Western Blot showed there was a huge difference in ATF5prote in expression (P<0.05).5、HE dyeing observation showed that the split phaseof specific transfection group decreased significantly, the apoptosis phenomenon is obvious.Conclusion:Silence ATF5restrains transplantation tumor growth in human epithelial ovarian ca ncer nude mouse, it may become a new target of ovarian cancer gene therapy.