摘要
目的观察肝癌细胞株HepG2自身分泌的白细胞介素-6（IL-6）对其增殖能力和细胞周期的影响。方法RNA干扰技术沉默IL-6基因，同时设空白对照组（不加转染试剂）、正常对照组（转染空脂质体）、无义对照组[转染无义对照小干扰RNA（siRNA）]，实验对照组（转染沉默IL-6的siRNA）。噻唑蓝（MTY）比色法检测细胞的增殖能力，用流式细胞技术检测细胞的生长周期。结果逆转录-聚合酶链反应（RT—PCR）检测各组IL-6mRNA的表达量分别为104．00±10．71、309．00±15．19、162．50±9．85、26．75±7．54；MTT法检测各组细胞增殖率分别为100．00％、160．72％、130．39％、55．54％；流式细胞技术检测空白对照组，正常对照组，无义对照组，实验对照组不同细胞周期时细胞比例分别为：G0／G1期分别为（68．09±0．98）％、（63．28±0．92）％、（67．01±0．93）％、（73．43±1．16）％；S期分别为（23．04±0．99）％、（32．32±0．83）％、（25．11±1．24）％、（20．34±0．20）％；S＋G2／M期分别为（31．91±0．98）％、（36．72±0．92）％、（32．99±0．93）％、（26．57±1．16）％。结论mRNA的RNA干扰技术能有效的抑制IL-6mRNA在肝癌细胞株HepG2中的表达，IL-6被沉默能抑制肝癌细胞株HepG2的增殖，使其生长周期延长；肝癌细胞株HepG2IL-6表达增高能促进其增殖，使其生长周期缩短。
Objective study the effect of self-secreted interleukin-6 （IL-6） of proliferation and cell cycle of liver cancer cell line HepG2. Methods The interleukin-6 was made to be silence by using RNA interference technology, and then the control groups were established accordindy by using small inter- fering RNA （siRNA） with insignificant order, liposome only and no transfection and transfected IL-6- RNA. Detect the proliferation and cell cycle by using methyl thiazol tetrazolium （MTT） and flow cytometry technology. Results The expression of IL-6 mRNA were 104. 00±10. 71, 309. 00±15.19, 162. 50 ±9.85, 26. 75±7.54; The cell proliferation rate were 100. 00%, 160. 72%, 130. 39%, 55.54% by using MTT; the cell proportion in different cell circles was detected by using Flow cytometry technology in different teams as follows ： G0/G1 phase account for （ 68.09±0. 98 ） % , （ 63.28±0. 92 ） % , （ 67.01±0.93）%, （73.43 ＋1.16）% ; S phase account for （23.04±0.99）%, （32.32±0.83）%, （25.11±1.24）%, （20.34 ＋0.20）%; S ＋G2/M phase account for （31.91±0.98）%, （36.72±0.92）%, （ 32. 99±0. 93 ） % , （ 26. 57±1.16） %. Conclusion It can supress the expression of IL-6 mRNA in liv- er cancer cell HepG2 hy using RNAi interference technique; it can supress the proliferation and extend the cell circle of liver cancer cell HepG2 when IL-6was silence ; its proliferation was promoted and cell circle was shorten when it secreted higher interleukin-6 in liver cancer cell line HepG2.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第6期1232-1234,共3页
Chinese Journal of Experimental Surgery
