天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (10): 1009-1012.doi: 10.11958/20170746

• 细胞与分子生物学 •    下一篇

CTCF 过表达对乳腺癌细胞凋亡因子 Bax 和 Bcl-2 表达的影响

张晓宇 1，康晓宁 2，刘伟 1，刘志明 1，王鹏 1，王遵义 1△   

  1. 1 河北省沧州市中心医院肿瘤外三科（邮编 061001），2 超声二科
  • 收稿日期:2017-06-28 修回日期:2017-07-12 出版日期:2017-10-15 发布日期:2017-10-13
  • 通讯作者: 张晓宇 E-mail:93956466@qq.com
  • 作者简介:张晓宇（1983），男，主治医师，硕士，主要从事乳腺癌综合治疗的研究

Effects of overexpression of CTCF on apoptosis factors Bax and Bcl-2 in breast cancer cells

ZHANG Xiao-yu1, KANG Xiao-ning2, LIU Wei1, LIU Zhi-ming1, WANG Peng1, WANG Zun-yi1△   

  1. 1 Department of Oncological Surgery-3, 2 Department of Ultrasound-2，Cangzhou Central Hospital, Hebei 061001, China
  • Received:2017-06-28 Revised:2017-07-12 Published:2017-10-15 Online:2017-10-13

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摘要： 目的 探讨 CTCF 过表达对人乳腺癌细胞系 MDA-MB-231 中细胞凋亡因子 Bax 和 Bcl-2 表达的影响。 方法 用逆转录聚合酶链式反应（RT-PCR）检测人乳腺癌细胞系 MDA-MB-231 中 CTCF、Bax 和 Bcl-2 的表达。构 建 CTCF/pEGFP-N1 过表达载体，用慢病毒转染法分别将 CTCF/pEGFP-N1 和空载质粒转入 MDA-MB-231 中，记作 CTCF 组和对照组。经 RT-PCR 鉴定 CTCF 成功转染 MDA-MB-231 后，采用实时定量 PCR（Q-PCR）检测 CTCF 组 和对照组 MDA-MB-231 中 Bax 和 Bcl-2 的 mRNA 表达，Western blot 和酶联免疫吸附试验（ELISA）检测 Bax 和 Bcl- 2 的蛋白表达。结果 MDA-MB-231 中不表达 CTCF，表达 Bax 和 Bcl-2。Q-PCR 结果显示，CTCF 组、对照组中 Bax 的 mRNA 表达水平分别为 4.63±1.08 和 2.27±0.16，2 组间差异有统计学意义（t=27.50，P＜0.05）；Bcl-2 的 mRNA 表 达水平分别为 1.39±0.14 和 3.56±0.97，2 组间差异有统计学意义（t=39.00，P＜0.05）。Western blot 结果显示，CTCF 组 中 Bax 蛋白表达水平高于对照组，Bcl-2 蛋白水平低于对照组。ELISA 结果显示，CTCF 组和对照组中 Bax 蛋白表达 水平分别为 15.25±2.17 和 6.24±1.78，2 组间差异有统计学意义（t=26.84，P＜0.05）；Bcl-2 蛋白表达水平分别为 4.59± 0.97 和 10.68±1.93，2 组间差异有统计学意义（t=21.72，P＜0.05）。结论 CTCF 过表达可促进乳腺癌细胞中凋亡因 子的表达，抑制抗凋亡因子的表达。

关键词: 乳腺肿瘤, bcl-2 相关 X 蛋白质, 基因, bcl-2, CTCF, 凋亡因子

Abstract: Objective To investigate the effect of the over- expressed CTCF on apoptosis factors Bax and Bcl- 2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF，Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDAMB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl- 2 were 4.59±0.97 and 10.68± 1.93, and there was significant difference between the two groups (t=21.72, P＜0.05). Conclusion The over- expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.

Key words: breast neoplasms, bcl-2-associated X protein, genes, bcl-2, CTCF, apoptosis factors