XPD Functions as a Tumor Suppressor and Dysregulates Autophagy in Cultured HepG2 Cells.
XPD Functions as a Tumor Suppressor and Dysregulates Autophagy in Cultured HepG2 Cells.
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XPD 在培养的 HepG2 细胞中发挥肿瘤抑制作用并调节自噬
DOI：
10.12659/msm.894303
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发表时间：
2015-05-29
期刊：
影响因子：
--
通讯作者：
Zhang JX
中科院分区：
文献类型：
--
作者：
Zheng JF;Li LL;Lu J;Yan K;Guo WH;Zhang JX
背景最近的临床研究表明，与核苷酸切除修复相关的关键修复基因--着色性干皮病D组(XPD)基因的多态与肝细胞癌的风险增加有关。然而，XPD在培养的肝癌细胞中的表达对细胞的影响在很大程度上仍不清楚。因此，本研究的目的是研究XPD基因的表达对肝癌细胞株HepG2的体外细胞效应。材料/方法将pEGFP-N_2/XPD质粒(XPD)、pEGFP-N_2质粒组(N_2)、脂质体™2000(脂质体)和未转染组(CON)分别转染人肝癌细胞株HepG2细胞。采用细胞增殖实验、Annexin V-APC细胞凋亡实验、集落形成实验、划痕迁移实验、Transwell迁移实验和Western blotting检测自噬蛋白Lc3和p62的表达。结果XPD表达显著抑制HepG2细胞增殖(p<；0.05)，显著促进细胞凋亡(p<；0.05)，显著抑制HepG2细胞集落形成(p<；0.05)，显著降低细胞迁移能力(p<；0.05)，显著降低细胞侵袭能力(p<；0.05)。Western blotting结果显示，XPD表达显著增加Lc3的表达(p<；0.05)，显著降低p62的表达(p<；0.05)。结论XPD的表达可抑制体外培养的HepG2细胞中自噬蛋白的降解，抑制肿瘤生长。需要进一步的体内临床前研究和临床试验来验证XPD作为一种肿瘤抑制基因疗法的潜力。
Background Recent clinical studies have linked polymorphisms in the xeroderma pigmentosum group D (XPD) gene, a key repair gene involved in nucleotide excision repair, to increased risk of hepatocellular carcinoma (HCC). However, the cellular effects of XPD expression in cultured HCC cells remain largely uncharacterized. Therefore, the aim of this study was to characterize the in vitro cellular effects of XPD expression on the HCC cell line HepG2. Material/Methods HepG2 cells were transfected as follows to create four experimental groups: pEGFP-N2/XPD plasmid (XPD) group, EGFP-N2 plasmid (N2) control group, lipofectamine™ 2000 (lipid) control group, and non-transfected (CON) control group. An MTT cell proliferation assay, Annexin V-APC apoptosis assay, colony formation assay, scratch wound migration assay, Transwell migration assay, and Western blotting of the autophagic proteins LC3 and p62 were conducted. Results XPD expression significantly inhibited HepG2 cell proliferation (p<0.05), significantly promoted HepG2 cell apoptosis (p<0.05), significantly inhibited HepG2 colony formation (p<0.05), significantly decreased HepG2 cells’ migratory ability (p<0.05), and significantly lowered HepG2 cells’ invasive capacity (p<0.05). Western blotting showed that XPD expression significantly increased LC3 expression (p<0.05) and significantly reduced p62 expression (p<0.05). Conclusions XPD expression serves as a tumor suppressor and dysregulates autophagic protein degradation in HepG2 cells in vitro. Further in vivo pre-clinical studies and clinical trials are needed to validate XPD’s potential as a tumor-suppressive gene therapy.