[c-myc反义寡核苷酸转染诱导HeLa细胞系凋亡]

[Inducing apoptosis of HeLa cell lines by transfection of c-myc antisense oligonucleotides].

作者信息

Li Rong-rong, Ma Dao-Xin, Cheng Yu-feng

机构信息

Department of Radiotherapy, Qilu Hospital, Shandong University, Jinan 250012, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Jan;37(1):19-22.

PMID:16468633

Abstract

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.

摘要

目的

通过转染c-myc反义寡核苷酸研究其对HeLa细胞凋亡的影响，寻求提高肿瘤细胞放射敏感性的新方法。

方法

采用免疫组织化学染色和逆转录-聚合酶链反应（RT-PCR）鉴定反义寡脱氧核苷酸（ASODN）的作用。通过吉姆萨染色、流式细胞术（FCM）和DNA梯状条带检测HeLa细胞的凋亡情况。

结果

转染ASODN（c-myc）后，c-myc mRNA和C-MYC蛋白表达水平均降低（P < 0.05）。吉姆萨染色显示c-myc ASODN转染细胞出现凋亡小体。DNA梯状条带呈现典型的凋亡条带。转染c-myc ASODN的HeLa细胞凋亡率为（10.29±0.66）%，转染c-myc ASODN联合照射组为（16.83±0.57）%，均显著高于对照组（1.79±0.19）%（P < 0.05）。