不同剂量60Co-γ射线对TM3细胞的损伤效应
| 作者: |
1,2张烨敏,
3欧俊,
2,4焦婷婷,
1邹冬芳,
2惠宁,
1孙健
1 解放军第85医院妇产科，上海 200052 2 第二军医大学附属长海医院妇产科，上海 200433 3 上海交通大学附属仁济医院生殖中心，上海 200135 4 解放军北京军区总医院生殖中心，北京 100700 |
| 通讯: |
惠宁
Email: huin1956@163.com 孙健 Email: 11sunjian@sina.com |
| DOI: | 10.3978/j.issn.2095-6959.2016.03.009 |
| 基金: | 军队基金 |
摘要
目的：对不同剂量60Co-γ射线下TM3细胞的损伤进行细胞生物学行为、转录层面的分析。方法：TM3细胞分4组，包括3个60Co-γ射线照射组(3、6、9 Gy)及对照组(不照射)。于24、48、72 h观察各组细胞凋亡情况(流式细胞法)、氧化损伤(ELISA)、睾酮合成关键因子(StAR、P450scc、P450c17及3β-HSD)的mRNA表达(实时定量PCR法)。于24、48、72、96、120、144 h，观察细胞增殖情况(MTS法)。结果：第24、72 h，各照射组细胞凋亡率均高于对照组(P<0.008)；MTS实验中，第144、120 h，各照射组OD490值均低于对照组(P<0.008)；第24，48，72 h，3 Gy组8-OHdG浓度与对照组相比无差异(P>0.008)，而9 Gy组高于对照组(P<0.008)；在72 h，各照射组StAR、P450scc、3β-HSD mRNA表达均低于对照组(P<0.008)，而P450c17的表达在3 Gy组中无显著改变，在6 Gy组中高于对照组，在9 Gy组中低于对照组。结论：60Co-γ射线使TM3细胞凋亡增加、细胞增殖能力下降。6 Gy、9 Gy照射可以导致TM3细胞DNA的氧化损伤。辐照导致TM3细胞中StAR、P450scc、3β-HSD mRNA表达下降。6 Gy的照射使P450c17 mRNA表达上升，9 Gy使之下降。
关键词：
60Co-γ射线
TM3细胞
细胞凋亡
细胞增殖
睾酮合成关键因子
Effect of different doses of 60Co γ- radiation on TM3 cells
CorrespondingAuthor: HUI Ning Email: huin1956@163.com
DOI: 10.3978/j.issn.2095-6959.2016.03.009
Abstract
Objective: To analyze the impairment of TM3 cells induced by 60Co γ radiation on cytobiological behaviors and transcriptional change. Methods: TM3 cells were divided into four groups, including three groups exposed to different doses of 60Co γ radiation (3, 6, 9 Gy) and one control group without exposure. At 24, 48, 72 h, cell apoptotic rate of different groups was measured by flowcytometry; oxidative damage was measured by ELISA; mRNA expression of the key factors in the synthesis of testosterone (StAR, P450Scc, P450c17, 3β-HSD) was measured by qPCR. At 24, 48, 72, 96, 120, 144 h, cell proliferation was measured by MTS. Results: At 24, 72 h, apoptotic rate of all exposed groups was obviously higher than that of the control group (P<0.008); in MTS assay, at 144, 120 h, the OD490 value of all exposed groups was lower than that of the control group (P<0.008). At 24, 48, 72 h, no significant difference of 8-OHdG level was observed between 3 Gy group and control group (P>0.008); 8-OHdG level of the 9 Gy group was higher than that of the control group (P<0.008). At 72 h, the mRNA expression of StAR, P450scc and P450scc in all exposed groups was down-regulated, as compared to the control group (P<0.008). However, the mRNA expression of P450c17 in 3 Gy group had no difference from that of the control group. It was up-regulated in 6 Gy Group and down-regulated in 9 Gy Group. Conclusion: 60Co γ radiation induced cell apoptosis and decreased cell proliferation in TM3 cells. A single dose of 6 or 9 Gy caused oxidative damage in TM3 cells. Radiation down-regulated the mRNA expression of the key factors of testosterone synthesis, including StAR, P450scc and 3β-HSD. 6 Gy radiation increased the P450c17 expression level, while 9 Gy radiation decreased it.