宫颈癌侧群细胞化疗耐受性的体外研究

The study on drug resistance of side population cells of cervical carcinoma in vitro

目的:观察化疗药物干预对人宫颈癌细胞系Hela侧群细胞和非侧群细胞中ABCG2、ABCC2基因表达变化的影响,探讨两群细胞化疗耐受性的差异。方法:Hoechst 33342免疫荧光染色与流式细胞术(FACS)相结合分选宫颈癌细胞系Hela中的侧群细胞和非侧群细胞。给予顺铂干预,利用RT-PCR观察两群细胞化疗前后ABCG2、ABCC2基因的表达差异,以及细胞凋亡情况。结果:与非侧群细胞相比,侧群细胞ABCG2、ABCC2基因表达更高,但差异不明显;顺铂干预后,两组细胞ABCG2、ABCC2基因表达均有升高,组间比较侧群细胞中ABCG2、ABCC2明显高于非侧群细胞,差异有统计学意义;侧群细胞对于IC50附近两个顺铂浓度0.95μg/ml和1.9μg/ml的耐受能力较好,细胞没有因药物的加入而大量凋亡,细胞凋亡率反而略有下降。非侧群细胞对于IC50附近0.95μg/ml和1.9μg/ml顺铂浓度的耐受能力较差,细胞大量凋亡。结论:来源于宫颈癌Hela细胞中的侧群细胞相对于非侧群细胞顺铂化疗耐受性更强,这种化疗耐受可能与侧群细胞ABCG2、ABCC2的高表达抑制细胞凋亡有关。

Objective： To observe the expression of ABCG2 and ABCC2 in side population cells（ SPCs） and non-side population cells（ NSPCs） originated from Hela clone of cervical carcinoma which were treated by cisplatin（ DDP）and to explore the difference of drug resistance between the two cell groups. Methods： SPCs and NSPCs were screened from Hela clone of human cervical carcinoma by using Hoechst 33342- stain method and flow cytometry. The expression of ABCG2 and ABCC2 in SPCs and NSPCs were detected by Real- Time PCR technology,and the ratio of apoptosis in the two group cells were calculated. Results： Expression of ABCG2 and ABCC2 in SPCs was higher than NSPCs,but there was no significant difference between the two groups（ P〉0. 05）. Expression of ABCG2 and ABCC2 rise in each group when the cells was treated with DDP,and there had significant difference between the two groups（ P〈0. 05）. The apoptosis ratio of SPCs basesd on IC50 values had almost not changed when the cells were treated with DDP,especially on the concentration of 0. 95μg / ml and 1. 9μg / ml,while there had lots of apoptosis in NSPCs,in particular on the concentration of 0. 95μg / ml and 1. 9μg / ml of DDP. Conclusion： SPCs which were separated from Hela cell line of human cervical carcinoma has stronger drug resistance ability than NSPCs deriving from the same cell clone. The rise of ABCG2 and ABCC2 expression may responsible for the drug resistance in SPCs,basing on the cell apoptosis number decreased.