钙网蛋白
基因敲除
转染
细胞凋亡
细胞内
脂质体
小干扰RNA
分子生物学
肝星状细胞
基因沉默
膜联蛋白
细胞生物学
生物
流式细胞术
化学
内质网
基因
生物化学
内分泌学
重组DNA
载体(分子生物学)
作者
Linyu Dai,Luguang Wang,Haiying Liu,Yonghong Xiao
出处
期刊:Chinese journal of cellular and molecular immunology
日期:2021-06-01
卷期号:37 (6): 501-506
被引量:3
摘要
Objective To investigate the effect of silencing calreticulin (CALR) gene on the apoptosis and intracellular Ca2+ concentration in HSC-LX2 human hepatic stellate cells. Methods Small interfering RNA (siRNA) targeting CALR was designed and transfected into HSC-LX2 cells by lipofectamine transfection, and then the cells with CALR knockdown were screened out. Apoptosis was detected by flow cytometry combined with annexin V-FITC/PI labeling. The intracellular Ca2+ concentration which was loaded with Fluo3-AM (calcium ion fluorescent probe) was observed by laser confocal microscope. The mRNA and protein levels of CALR, Bcl2 and BAX were detected by reverse-transcription PCR and Western blotting. Results Knockdown of CALR led to the increase of intracellular Ca2+ concentration, the increased apoptosis of HSC-LX2 cells, the up-regulation of BAX expression, down-regulation of Bcl2 and the obvious raise of BAX/Bcl2 ratio. Conclusion Knockdown of CALR can increase intracellular Ca2+ concentration, up-regulate the ratio of BAX/Bcl2 and promote the apoptosis of HSC-LX2 cells.
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