脂质体介导c-myc反义寡核苷酸诱导HeLa细胞凋亡的实验研究

目的体外研究C-MYC反义寡核苷酸(ASODN)对人宫颈癌HELA细胞凋亡的影响,寻求提高肿瘤细胞放射敏感性的方法。方法免疫组化、流式细胞术、半定量RT-PCR法鉴定ASODN的有效性;GIEMSA染色、流式细胞术检测凋亡指数(AI)、DNA LADDER检测细胞凋亡。结果转染后,免疫组化结果显示C-MYC蛋白表达降低;半定量RT-PCR结果显示C-MYC MRNA表达较空白对照组及阴性对照组明显减弱(P<0.05)。GIEMSA染色可见凋亡小体;流式细胞术结果显示转染后凋亡指数为(10.29±0.66)%,转染ASODN(C-MYC)联合放射的凋亡指数为(16.83±0.57)%,均明显高于空白对照组(1.79±0.19)%(P<0.05);DNA LADDER呈现具有凋亡特征的梯带。结论本实验所设计的ASODN能有效地抑制C-MYC基因的表达;转染ASODN(C-MYC)能够显著增加HELA细胞凋亡,从而提高肿瘤细胞的放射敏感性。

Inducing Apoptosis of HeLa Cell Lines by Transfection of c-myc Antisense Oligonucleotides

OBJECTIVE: To investigate the effect on apoptosis of HeLa cells by transfecting c-myc antisense oligonucleotides and seek a new method to enhance radiosensitivity of tumor cells. METHODS: Immunohistochemistry staining and RT-PCR were conducted to identify the effect of ASODN. Giemsa staining, flow cytometry (FCM) and DNA ladder were performed to detect the apoptosis of HeLa cells. RESULTS: After transfection of ASODN (c-myc), the levels of both c-myc mRNA and C-MYC protein expression were reduced (P < 0.05). Giemsa staining demonstrated apoptosis-bodies on the c-myc ASODN-transfected cells. DNA ladder presented the typical ladder of apoptosis. The apoptosis rate of HeLa cells transfected with c-myc ASODN was (10.29 +/- 0.66)%, transfected c-myc ASODN combined irradiation was (16.83 +/- 0.57)%, which were much higher than that of control group (1.79 +/- 0.19)% (P < 0.05). CONCLUSION: The c-myc ASODN could effectively down-regulate the expression of c-myc; furthermore, transfection of c-myc ASODN could more effectively induce apoptosis of HeLa cells which subsequently enhanced radiosensitivity of HeLa cells.