慢病毒短发夹RNA介导的多形性腺瘤基因样蛋白2沉默对肝癌细胞MHCC97-L增殖、迁移及侵袭的影响#br#

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doi:10.16098/j.issn.0529-1356.2022.01.010

摘要/Abstract

摘要：

目的探讨慢病毒短发夹RNA（shRNA）介导的多形性腺瘤基因样蛋白2（PLAGL2）沉默对肝癌细胞恶性行为的影响及其机制。方法 Real-time PCR与Western blotting分别检测肝癌组织及癌旁组织中PLAGL2的表达水平；体外培养肝癌细胞MHCC97-L，构建慢病毒载体质粒PLAGL2-shRNA与对照NC-shRNA，转染MHCC97-L细胞，采用嘌呤霉素筛选稳转株；CCK-8、Transwell小室实验检测沉默PLAGL2后MHCC97-L细胞的增殖活力以及迁移和侵袭数量；Western blotting检测p-PI3K和p-Akt蛋白表达。用PI3K/Akt信号通路激活剂处理MHCC97-L细胞，检测细胞增殖、迁移及侵袭能力。结果肝癌组织中PLAGL2 表达水平显著升高（P＜0.05）；9株MHCC97-L细胞转染PLAGL2-shRNA可明显降低PLAGL2表达水平，同时，细胞增殖、迁移、侵袭能力减弱（P＜0.05），p-PI3K、p-Akt 表达受抑制（P＜0.05）；PI3K/Akt激活剂可明显逆转上述现象。结论慢病毒shRNA载体介导的PLAGL2沉默，可显著抑制肝癌细胞恶性行为，其作用机制可能与抑制PI3K/Akt信号通路的激活有关。

关键词: 慢病毒, 短发夹RNA, 多形性腺瘤基因样蛋白2, 肝癌, 磷脂酰肌醇-3激酶/蛋白酶B信号通路, 免疫印迹法, 人

Abstract:

Objective To investigate the effect of short hairpin RNA(shRNA)-mediated pleomorphic adenoma gene like-2 (PALAG2) silencing on the malignant behavior of hepatocellular carcinoma cells and its mechanism. Methods Real-time PCR and Western blotting were used to detect the expression level of PLAGL2 in liver cancer tissues and adjacent tissues. Hepatoma cells MHCC97-L were cultured in vitro, the lentiviral vector plasmid PLAGL2-shRNA and control NC-shRNA were constructed, transfected into MHCC97-L cells, and stable transfected strains were selected with puromycin. CCK-8 and Transwell chamber assay detected the proliferation activity and the number of migration and invasion of MHCC97-L cells after silencing PLAGL2. Western blotting was used to detect the expression of p-PI3K and p-Akt proteins. The PI3K/Akt signaling pathway activator was used to treat MHCC97-L cells to detect cell proliferation, migration and invasion. Results The expression of PLAGL2 was significantly increased in liver cancer tissue (P＜0.05). Transfection of 9 strains of MHCC97-L cells with PLAGL2-shRNA could significantly reduce the expression level of PLAGL2, and the ability of proliferation, migration, and invasion of MHCC97-L cells was also weakened (P<0.05), and the expression levels of p-PI3K, and p-Akt were inhibited (P<0.05), PI3K/Akt activator could obviously reverse the above phenomenon. Conclusion shRNA lentiviral vector pathway can effectively silence the expression of PLAGL2 gene in hepatocarcinoma cells. Silencing of PLAGL2 can significantly inhibit the malignant behavior of proliferation, migration and invasion of hepatocarcinoma cells, and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway activation.

Key words: Lentivirus, Short hairpin RNA, Pleomorphic adenoma gene like-2, Hepatocellular carcinoma, Phosphatidylinositol 3-kinase/protein kinase B signaling pathway, Western blotting, Human

中图分类号:

R735.7

赵克昌王伟杨金煜冯鹏才吴世乐 . 慢病毒短发夹RNA介导的多形性腺瘤基因样蛋白2沉默对肝癌细胞MHCC97-L增殖、迁移及侵袭的影响#br#

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[J]. 解剖学报, 2022, 53(1): 75-81.

ZHAO Ke-chang WANG Wei YANG Jin-yu FENG Peng-cai WU Shi-le. Effect of short hairpin RNA lentivirus-mediated silencing of pleomorphic adenoma gene like-2 on proliferation, migration and invasion of hepatocellular carcinoma cells[J]. Acta Anatomica Sinica, 2022, 53(1): 75-81.

参考文献

［1］ Zhou RJ, Lv HZ. Knockdown of ACTA-AS1 promotes liver cancer cell proliferation, migration and invasion［J］. Mol Med Rep, 2019, 19(3): 2263-2270.

［2］ Lin Q, Ling YB, Chen JW, et al. Circular RNA circCDK13 suppresses cell proliferation, migration and invasion by modulating the JAK/STAT and PI3K/AKT pathways in liver cancer［J］. Int J Oncol, 2018, 53(1): 246-256.

［3］ Yue X, Cui Y, You Q,et al. MicroRNA-124 negatively regulates chloride intracellular channel 1 to suppress the migration and invasion of liver cancer cells［J］. Oncol Rep, 2019, 42(4): 1380-1390.

［4］ Yang C, Xu Y, Cheng F, et al. miR-1301 inhibits hepatocellular carcinoma cell migration, invasion, and angiogenesis by decreasing Wnt/β-catenin signaling through targeting BCL9［J］. Cell Death Dis, 2017, 8(8): e2999-e3009.

［5］ Qu G, Xu Y, Wan SP, et al. Expression of PLAGL2 in bladder urothelial carcinoma and its relationship to lymph node metastasis and survival［J］. Sci Rep, 2018, 8(1): 6044-6054.

［6］ Wang XF, Lu YP. Expression of pleomorphic adenoma gene like-2 in gastric carcinoma and its possible mechanism on invasion and epithelial-mesenchymal transition of gastric carcinoma［J］. Chinese Journal of Experimental Surgery, 2019, 36(3): 432-435. (in Chinese)

王啸飞, 路夷平. 多形性腺瘤基因样因子2在胃癌组织中的表达及其影响胃癌细胞侵袭、上皮-间充质转化的机制［J］. 中华实验外科杂志, 2019, 36(3): 432-435.

［7］ Hu W, Zheng S, Guo H, et al. PLAGL2-EGFR-HIF-1/2α signaling loop promotes HCC progression and Erlotinib insensitivity［J］. Hepatology, 2021, 73(2):674-691.

［8］ Xu Zh, Zhang M, Zhang ZhL, et al. Effects of ADAMTS9 gene combined with demethoxycurcumin on invasion and migration of hepatocellular carcinoma cells and PI3K/PTEN/Akt signal［J］. Chinese Journal of Gerontology, 2019, 39(11): 2731-2735. (in Chinese)

徐卓, 张萌, 张志磊, 等. ADAMTS9基因联合去甲氧基姜黄素对肝癌细胞侵袭迁移及PI3K/PTEN/Akt信号的影响［J］. 中国老年学杂志, 2019, 39(11): 273-2735.

［9］ Li WF, Dai H, Ou Q, et al. Overexpression of microRNA-30a-5p inhibits liver cancer cell proliferation and induces apoptosis by targeting MTDH/PTEN/Akt pathway［J］. Tumor Biol, 2016, 37(5): 5885-5895.

［10］ Li Y, Cai Y, Chen H, et al. Clinical significance and association of GSTP1 hypermethylation with hepatocellular carcinoma: a meta-analysis［J］. J Cancer Res Ther, 2018, 14(Suppl):S486-S489.

［11］ Li GP, Liu LX, Pu ZJ, et al. Effects of human hypoxia inducible factor-1 alpha gene silencing by lentiviral-mediated shRNA on proliferation and apoptosis of HepG2 cells under hypoxia［J］. Medical Journal of Wuhan University, 2019, 40(1): 27-32. (in Chinese)

李国平, 刘丽璇, 蒲泽锦, 等. 慢病毒介导HIF-1α沉默对缺氧肝癌细胞HepG2增殖和凋亡的影响［J］. 武汉大学学报(医学版), 2019, 40(1): 27-32.

［12］ He P, Zheng L, Wang J, et al. The research progress in PLAG gene family［J］. Genomics and Applied Biology, 2019, 38(3): 1065-1069. (in Chinese)

何盼, 郑立, 王健, 等. PLAG基因家族研究进展［J］. 基因组学与应用生物学, 2019, 38(3): 1065-1069

［13］ Wang QY, Li XX, Zhu JY. Expression and clinical significance of miR-449a and PLAGL2 in ovarian cancer［J］. Journal of Medical Research, 2019, 48(3): 105-110. (in Chinese)

王秋宇, 李晓翔, 朱军义. MiR-449a和PLAGL2在卵巢癌中的表达关系及临床意义［J］. 医学研究杂志, 2019, 48(3): 105-110.

［14］ Qu GY, Xu Y, Liu JG, et al. Expression and clinical significance of PLAGL2 in prostate cancer［J］. Chinese Journal of Cancer Prevention and Treatment, 2018, 25(20): 1446-1451. (in Chinese)

瞿根义, 徐勇, 刘劲戈, 等. 前列腺癌组织PLAGL2表达临床意义探讨［J］. 中华肿瘤防治杂志, 2018, 25(20): 1446-1451.

［15］ Gao J, Wang R, Yang QL, et al. The effect of oxaliplatin on cell cycle of hepatoma cell HepG_2［J］. Journal of Zhejiang University (Medical Edition), 2013, 42(4): 437-442. (in Chinese)

高洁, 汪蕊, 杨清玲, 等. 奥沙利铂对肝癌细胞HepG_2细胞周期的影响［J］. 浙江大学学报(医学版), 2013, 42(4): 437-442.

［16］ Wang XJ, Zhao YN, Li X. Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells［J］. Acta Anatomica Sinica, 2018, 49(3): 342-346. (in Chinese)

王小杰, 赵依纳, 李欣. 敲低膜联蛋白A5对人胃癌MGC-803和MKN-45细胞周期的影响［J］. 解剖学报, 2018, 49(3): 342-346.

［17］ Yang J, Tan GH, Chen JZ. Lentivirus-mediated shRNA interference of Cx26 suppresses epithelial mesenchymal transi-tion and invasion of highly invasive hepatocellular carcinoma cells in vitro［J］. Journal of Southern Medical University, 2014, 34(12): 1743-1747. (in Chinese)

阳洁, 覃贵慧, 陈军泽. 慢病毒介导shRNA靶向下调Cx26表达对人高侵袭性肝癌细胞上皮间质转化及侵袭的影响［J］. 南方医科大学学报, 2014, 34(12): 1743-1747.

［18］ Tian LF, Zhang Q, Wang BT, et al. Mechanism of sanguinarine in inducing apoptosis in murine pancreatic cancer cells by inhibiting PI3K/Akt signaling pathway［J］. Chinese Journal of Experimental Traditional Medical Formulae, 2018, 24(19): 166-171. (in Chinese)

田龙夫, 张琦, 王波涛, 等. 血根碱通过调控PI3K/Akt信号通路诱导胰腺癌细胞凋亡的机制［J］. 中国实验方剂学杂志, 2018, 24(19): 166-171.

［19］ Fu Y, Hong ZhQ, Li N, et al. Research on the effect of Pien Tze Huang on human osteosarcoma MG63 cell proliferation and PI3K/Akt signal pathway［J］. Journal of Traditional Chinese Medicine Pharmacy, 2016, 31(11): 4675-4677. (in Chinese)

伏勇, 洪振强, 李楠, 等. 片仔癀对人骨肉瘤MG63细胞增殖及PI3K/Akt信号通路的作用研究［J］. 中华中医药杂志, 2016, 31(11): 4675-4677.

［20］ Hu ZCh. Effects of juglandis immaturum exocarpium polysaccharide on proliferation, apoptosis and PI3K/Akt signaling pathway of human colon carcinoma cell line HCT-116［J］. Chinese Journal of Experimental Traditional Medical Formulae, 2016, 22(18): 136-139. (in Chinese)

胡泽成. 青龙衣多糖对结肠癌HCT-116细胞增殖、凋亡及PI3K/Akt信号通路的影响［J］. 中国实验方剂学杂志, 2016, 22(18): 136-139.

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慢病毒短发夹RNA介导的多形性腺瘤基因样蛋白2沉默对肝癌细胞MHCC97-L增殖、迁移及侵袭的影响#br#
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Effect of short hairpin RNA lentivirus-mediated silencing of pleomorphic adenoma gene like-2 on proliferation, migration and invasion of hepatocellular carcinoma cells

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