Zhang Lei, Gao Aijun, Peng Kaiyun
Department of Medical Laboratory Technology, Medical College, Yangzhou Polytechnic College, Yangzhou 225009, China.
Department of Medical Laboratory Technology, Medical College, Yangzhou Polytechnic College, Yangzhou 225009, China.
Arab J Gastroenterol. 2025 Aug;26(3):241-249. doi: 10.1016/j.ajg.2025.05.002. Epub 2025 Jun 28.
Sorafenib, as a novel multi-targeted oral tumor chemotherapeutic drug, has been found to exert an impact on the inhibition of cancer growth. Phosphatidylinositol-binding reticulin assembly protein interacting with mitotic regulatory factors (PIMREG) is strongly associated with oncology to drug resistance. However, how PIMREG modulates therapy tolerance to sorafenib in HCC and its potential regulatory mechanisms remain unclear. This study is abouta mechanistic approach to examine the action and mechanism of PIMREG in HCC-mediated sorafenib resistance.
The human hepatocellular carcinoma sensitive cell line Huh7 and drug-resistant cell line Huh7/SFB were used for the study, and different rates of PIMREG expansion in both cells were detected. Next, the study transfected PIMREG overexpression and interference vector into hepatoma cell line Huh7/SFB, and acted on the cells with solafenib exhibiting a concentration gradient. The growth inhibition rate and IC50 value of cells were detected by MTT method to determine the concentration and time of drug addition. Then, this study employed MTT, qRT-PCR, flow cytometry, and Western blot to assay the growth of these cells, which were induced through overexpression and disruption of PIMREG, in combination with sorafenib. The study also constructed an in vivo mouse tire sample test in order to investigate the influence of PIMREG upon the in vitro efficacy of sorafenib. In addition, the study used LY294002 inhibitors to explore the molecular mechanisms of PIMREG-mediated resistance to sorafenib in Huh7/SFB cells.
The expression level of PIMREG in cells of the Huh7/SFB resistant strain was clearly higher than that in cells of the sensitive strain Huh7. After transfection of sh-PIMREG, the IC50 value decreased significantly, while OE-PIMREG significantly increased the IC50 value of sorafinib. Compared with the control group, inhibition of cell proliferation by sorafenib was enhanced after interference with PIMREG, while the effect of overexpression of PIMREG was on the contrary. The efficacy of sorafenib was enhanced by knockout of PIMREG in living organisms. In addition, the PI3K/AKT signal pathway was necessary for PIMREG-induced sorafenib resistance. Subsequently, PIMREG regulated sorafenib-induced inhibition of the PI3K/AKT signaling pathway, and LY294002 blocked the signal pathway to reduce PIMREG-induced resistance.
All in all, an increase in HCC resistance to sorafenib via the PIMREG-mediated PI3K/AKT pathway suggests that PIMREG is a key tumor-associated gene with significant implications for sorafenib resistance in tumor cells.
索拉非尼作为一种新型的多靶点口服肿瘤化疗药物，已被发现对抑制癌症生长有影响。与有丝分裂调节因子相互作用的磷脂酰肌醇结合网硬蛋白组装蛋白（PIMREG）与肿瘤对药物的耐药性密切相关。然而，PIMREG如何调节肝癌对索拉非尼的治疗耐受性及其潜在的调控机制仍不清楚。本研究采用一种机制性方法来研究PIMREG在肝癌介导的索拉非尼耐药中的作用及机制。
本研究使用人肝癌敏感细胞系Huh7和耐药细胞系Huh7/SFB，检测两种细胞中不同比例的PIMREG扩增情况。接下来，将PIMREG过表达和干扰载体转染到肝癌细胞系Huh7/SFB中，并用呈浓度梯度的索拉非尼作用于细胞。采用MTT法检测细胞的生长抑制率和IC50值，以确定药物添加的浓度和时间。然后，本研究采用MTT、qRT-PCR、流式细胞术和蛋白质免疫印迹法检测这些通过PIMREG过表达和干扰诱导的细胞与索拉非尼联合作用后的生长情况。该研究还构建了体内小鼠肿瘤样本试验，以研究PIMREG对索拉非尼体外疗效的影响。此外，该研究使用LY294002抑制剂来探究PIMREG介导的Huh7/SFB细胞对索拉非尼耐药的分子机制。
耐药株Huh7/SFB细胞中PIMREG的表达水平明显高于敏感株Huh7细胞。转染sh-PIMREG后，IC50值显著降低，而OE-PIMREG则显著提高了索拉非尼的IC50值。与对照组相比，干扰PIMREG后索拉非尼对细胞增殖的抑制作用增强，而过表达PIMREG的效果则相反。在生物体中敲除PIMREG可增强索拉非尼的疗效。此外，PI3K/AKT信号通路是PIMREG诱导索拉非尼耐药所必需的。随后，PIMREG调节索拉非尼诱导的对PI3K/AKT信号通路的抑制，LY294002阻断该信号通路以降低PIMREG诱导的耐药性。
总之，肝癌通过PIMREG介导的PI3K/AKT途径对索拉非尼耐药性的增加表明，PIMREG是一个关键的肿瘤相关基因，对肿瘤细胞中索拉非尼耐药具有重要意义。
