【摘要】 背景:GATA5蛋白在多种癌症发生过程中被抑制表达,有研究显示GATA5降低表达或被甲基化后失去功能,不能有效促进干细胞分化,维持细胞的正常功能。宫颈癌HELA细胞表达GATA5蛋白,在宫颈癌中抑制GATA5表达是否可促进其恶性转化还是未知。目的:分析GATA5蛋白在宫颈癌HELA细胞中被进一步抑制表达后,其恶性转化情况及与癌干细胞形成相关因子sox2,c-myc及CD44的表达情况。方法:转入CMV-siRNA-gata5,使GATA5在HELA细胞中被进一步抑制表达后,(1)RT-PCR检测c-myc mRNA,sox2 mRNA的表达;(2)MTT法检测HELA细胞的增殖能力;(3)检测HELA细胞的集落形成和软琼脂单克隆形成能力;(4)Western blot检测细胞GATA5、c-myc、sox2蛋白表达变化;(5)激光共聚焦检测细胞的CD44蛋白定位及蛋白表达变化。结果与结论:用CMV-siRNA-gata5抑制HELA细胞中的GATA5表达后,HELA细胞的增殖能力、单克隆形成及集落形成能力均增强;同时发现其癌干细胞因子c-myc,sox2,CD44的表达增强。提示GATA5蛋白被抑制表达后,HELA细胞的恶行性为增加,其机制可能是GATA5被抑制后,促进了癌干细胞因子sox2,c-myc,CD44等的表达。更多还原

【Abstract】 BACKGROUND:GATA5, as a tumor suppressor gene, plays a role in inhibiting the development of cancer.GATA5 can be methylated in gastrointestinal tumors and then lose its function. GATA5 can be also methylated during hepatocarcinogenesis, and then cannot promote the differentiation of stem cells and maintain the normal function. Cervical cancer HELA cells express GATA5 protein, in which whether GATA5 plays a tumor suppressor function is still unknown.OBJECTIVE:To analyze the expression of sox2, c-myc and CD44 related to cancer stem cell formation after suppressing the expression of GATA5 in cervical cancer HELA cells.METHODS:Transfection of CMV-siRNA-gata5 further inhibited the expression of GATA5 in cervical cancer HELA cells. The expression of c-myc mRNA and sox2 mRNA were then detected by RT-PCR. The proliferation of HELA cells was detected by MTT. The colony formation and soft agar monoclonal formation of HELA cells were observed The expression of GATA5, c-myc and sox2 was detected by western blot. The expression and location of CD44 was detected by laser confocal microscope.RESULTS AND CONCLUSION:After suppressing the expression of GATA5 in cervical cancer HELA cells by CMV-siRNA-gata5, the proliferation, cell monoclonal formation and colony formation of HELA cells were enhanced The expression of c-myc, sox2 and CD44 was also strengthened. These findings indicate that the malignant activity of HELA cells is increased by inhibiting the GATA5 protein expression. The mechanism may be that inhibition of GATA5 promotes the expression of cancer stem cell factors sox2, c-myc, and CD44.更多还原