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shRNA 介导的热休克蛋白 70 基因沉默抑制人结肠癌生长。

ShRNA-mediated gene silencing of heat shock protein 70 inhibits human colon cancer growth.

机构信息

Department of General Surgery, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, P.R. China.

出版信息

Mol Med Rep. 2011 Sep-Oct;4(5):805-10. doi: 10.3892/mmr.2011.528. Epub 2011 Jul 1.

Abstract

Heat shock protein 70 (Hsp70), a chaperone involved in tumor progression, is overexpressed in various human tumors. However, its role in colon cancer progression is not completely understood. In the present study, two shRNA plasmid vectors against Hsp70 were constructed and stably transfected into the colon cancer cell line HT29 to determine the effect of Hsp70 on cell proliferation, cell cycle distribution and cell apoptosis in HT29 cells in vitro, and its effect on xenograft tumor growth and apoptosis in vivo. Cell proliferation was determined using MTT assay. The results revealed that Hsp70 silencing efficiently inhibited the growth of HT29 cells in culture, induced cell cycle arrest at the G1 phase, and significantly increased apoptosis. Moreover, stable clones from the Hsp70 shRNA-2 vector suppressed xenograft tumor growth and enhanced apoptosis in vivo compared with a mock and vector control group. In conclusion, specific Hsp70 shRNA silencing may inhibit colon cancer growth, indicating that Hsp70 silencing is a potential therapeutic strategy for the treatment of colon cancer.

摘要

热休克蛋白 70(Hsp70)是一种参与肿瘤进展的伴侣蛋白,在各种人类肿瘤中过度表达。然而,其在结肠癌进展中的作用尚不完全清楚。在本研究中,构建了针对 Hsp70 的两个 shRNA 质粒载体,并稳定转染至结肠癌 HT29 细胞系,以确定 Hsp70 对 HT29 细胞体外增殖、细胞周期分布和细胞凋亡的影响,以及其对体内异种移植肿瘤生长和凋亡的影响。通过 MTT 法测定细胞增殖。结果表明,Hsp70 沉默可有效抑制 HT29 细胞的体外生长,诱导细胞周期停滞在 G1 期,并显著增加细胞凋亡。此外,与 mock 和载体对照组相比,Hsp70 shRNA-2 载体的稳定克隆可抑制体内异种移植肿瘤的生长并增强凋亡。总之,特异性 Hsp70 shRNA 沉默可能抑制结肠癌的生长,表明 Hsp70 沉默是治疗结肠癌的一种潜在治疗策略。

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