基因特异性沉默对宫颈癌细胞增殖和凋亡的影响.docxVIP

AbstractoBJECTIVE Abstract oBJECTIVE To investigate the proliferative activity and apoptosis of HeLa cells after HPV 1 8 E6 was knocked down by specific synthetic siRNA． METHoDS The chemically synthetic siRNA targeting HPVl 8 E6 Was transfected into HeLa cells by Lipofectamine 2000，then the mRNA levels of HPV 1 8 E6 and hWAPL(human wings apart-like，hWAPL)were testeA by RT-PCR．MTT，immunocytochemistry，flow cytometry and transmission electron microscope were used to detect proliferation， expression of p53 protein，cell cycleapoptosis and variation of ultramicrostructure， respectively． RESUI』rS MTT results showed the decrease of proliferative activity in HeLa cells；RT-PCR results suggested lower expression of E6 and h WAPL rnRNA compared with control group，which is lowest at 48h after the transfeetion，while higher level of p53 protein expression Was detected by immunocytochemistry；flow cytometry revealed obvious G1 arrest in cell cycle and no significative apoptosis；mitochondria swelling and scKrcc apoptosis were observed by transmission electron microscope． CoNCLUSIoN The chemically synthetic siRNA could effectively silence the expression of E6， lower the level of hWAPL transcription and up-regulate the expression of p53 protein．Finally it induces some changes of HeLa cells，including decrease of proliferative activity，G1 arrest in cell cycle，variation ofultramicrostructure． Postgraduate student：Guang-Jie Yin(Gynecology and Obstetrics) Directed by Prof’··-。Yan-Kui Wang Bing Luo Key words：RNA interference；Human papillomavirus 18：Uterine Cervical Neoplasms；HeLa cells；Genes，WAPL，human 学位论文独创性声明、学位论文知识产权权属说明学位论文独创性声明、学位论文知识产权权属声明 学位论文独创性声明、学位论文知识产权权属说明 学位论文独创性声明、学位论文知识产权权属声明 学位论文独创性声明 本人声明，所呈交的学位论文系本人在导师指导下独立完成的研究成果。文 中依法引用他人的成果，均已做出明确标注或得到许可。论文内容未包含法律意义 上已属于他人的任何形式的研究成果，也不包含本人已用于其他学位申请的论文或 成果。 本人如违反上述声明，愿意承担由此引发的一切责任和后果。 论文作者签名： 耐治 日期：加，8年瑚阳 学位论文知识产权权属声明 本人在导师指导下所完成的学位论文及相关的职务作品，知识产权归属学 校。学校享有以任何方式发表、复制、公开阅览、借阅以及申请专利等权利。本人 离校后发表或使用学位论文或与该论文直接相关的学术论文或成果时，署名单位仍 然为青岛大学。