Effect of human protein kinase R-like endoplasmic reticulum kinase gene on the apoptosis of LO2 hepatocytes under endoplasmic reticulum stress

文献类型： 外文期刊

第一作者： Qin, Dongmei

作者： Qin, Dongmei;Li, Li;Zhang, Wei;Qin, Dongmei;Zhang, Yunsheng

作者机构：

关键词： Lentivirus;hepatocyte;protein kinase R-like endoplasmic reticulum kinase;gene modification

期刊名称：INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE （ 影响因子：0.166； 五年影响因子：0.621 ）

ISSN： 1940-5901

年卷期： 2016 年 9 卷 5 期

页码：

收录情况： SCI

摘要： Background: To evaluate the effects of recombinant lentiviral particles for human lentiviral vector carrying protein kinase R-like endoplasmic reticulum kinase (PERK) gene on endoplasmic reticulum (ER) stress-mediated apoptosis of human LO2 hepatocytes. Methods: 293T cells were co-transfected with recombinant lentiviral vector pWPT-GFP-PERK as well as packaging plasmids pMD2G and pSPAX2, giving a PERK-expressing lentivirus (LV-PERK). The supernatant was collected 48 h after transfection to infect LO2 hepatocytes. Infection efficiency was detected by observing GFP expression. The effects of LV-PERK on the apoptosis of LO2 cells under ER stress were assessed by flow cytometry. The expressions of apoptosis-related cleaved caspase-3 and CHOP proteins were detected by Western blotting. Results: Recombinant PERK lentivirus with titer of 4.2x10(8) efu/mL was constructed and packaged. Proportion of G1-phase cells in the tunicamycin (TM) + LV-PERK group was 53.49%, which was lower than those of TM (78.94%) and LV-GFP (65.73%) groups. The TM + LV-PERK group (32.45%) had significantly higher proportion of S-phase cells than those of TM (13.23%) and LV-GFP (17.79%) groups (P<0.05). The apoptotic rate of the TM + LV-PERK group (26.55%) significantly exceeded those of TM (12.59%) and LV-GFP (11.43%) groups (P<0.05). Western blotting results were consistent with those of flow cytometry. Conclusion: Under ER stress, LV-PERK promoted the proliferation and apoptosis of LO2 hepatocytes.

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