CPLX2在30例肝癌组织的表达及其对体外细胞增殖与侵袭的影响

doi:10.6040/j.issn.1671-7554.0.2020.0748

摘要/Abstract

摘要： 目的检测人复合素2(CPLX2)在肝癌组织中的表达,探讨其对肝癌细胞增殖与侵袭的影响。方法采用RT-qPCR和Western blotting方法检测30例配对肝癌组织和癌旁组织中CPLX2 mRNA和蛋白水平的表达。利用Lipofectamine 2000转染两条CPLX2小分子干扰RNA(siRNA)至肝癌Huh7细胞,RT-qPCR和Western blotting方法验证转染后的干扰效率。细胞实验分4组:空白组、对照siRNA组,CPLX2 siRNA1组和CPLX2 siRNA2组。噻唑蓝(MTT)法和Transwell实验检测细胞增殖和侵袭能力。采用成组t检验分析siRNA干扰效果和细胞侵袭能力的差异,双因素方差分析法两两多重比较各组细胞增殖能力的差异。结果 CPLX2在肝癌组织(22.69±14.78)中的表达高于癌旁组织(4.03±2.65),差异有统计学意义(t=5.941, P<0.001)。CPLX2 mRNA在两siRNA干扰组的表达量分别为0.34±0.02和0.48±0.01,均低于对照siRNA组(0.88±0.02)和空白组(1.00±0.05),差异有统计学意义(P<0.001),mRNA和蛋白水平均显示siRNA干扰效果较好。MTT实验证实CPLX2两siRNA干扰组的细胞增殖能力低于对照siRNA组(P<0.001)和空白组(P<0.001)。Transwell migration实验显示每个检测视野CPLX2 siRNA1组细胞的穿膜细胞数(37.0±2.0)和CPLX2 siRNA2组细胞的穿膜细胞数(46.3±2.5)低于空白组(88.0±2.0)和对照siRNA组(77.0±4.4)。Transwell invasion实验结果显示,每个检测视野CPLX2 siRNA1组细胞的穿膜细胞数(29.7±2.5)和CPLX2 siRNA2组细胞的穿膜细胞数(41.0±2.6)低于与空白组(74.7±3.1)和对照siRNA组(68.7±1.5),差异有统计学意义(P<0.001)。结论 CPLX2在肝癌组织中表达升高,下调其表达可抑制肝癌细胞Huh7的增殖和侵袭,CPLX2可能在促进肝癌的发生发展过程发挥重要作用。

关键词: 肝癌, 人复合素2, 增殖, 侵袭

Abstract: Objective To investigate the expression of Complexin 2(CPLX2)in human hepatocellular carcinoma(HCC)and its effects on the proliferation and invasion of Huh7 cells. Methods The mRNA and protein expressions of CPLX2 in 30 HCC tissues and adjacent tissues were detected with RT-qPCR and Western blotting. Two CPLX2 small interfering RNA(siRNA)were transfected into Huh7 cells mediated by Lipofectamine 2000. The transfection efficiency of siRNA was confirmed by RT-qPCR and Western blotting. The cells were divided into 4 groups: blank group, siRNA control group, CPLX2 siRNA group 1 and CPLX2 siRNA group 2. The cell proliferation and migration were determined with methyl thiazol tetrazolium(MTT)and Transwell assay. The siRNA interference effects and cell invasion were analyzed with group t test. The cell proliferation in each group was compared with two-way ANOVA. Results The expression of CLPX2 was higher in HCC tissues than in adjacent tissues ［(22.69±14.78) vs(4.03±2.65), t=5.941, P<0.001］. The mRNA expression of CPLX2 was(0.34±0.02)in CPLX2 siRNA group 1 and(0.48±0.01)in CPLX2 siRNA group 2, which was lower than that in the siRNA control group(0.88±0.02)and blank group(1.00±0.05), and the difference was statistically significant(P<0.05), showing good siRNA interference effects. MTT assay confirmed the cell proliferation was lower in CPLX2 siRNA groups 1 and 2 than in the control group(P<0.001)and blank group(P<0.001). Transwell migration showed the number of penetrating cells in each detection field was(37.0±2.0)in CPLX2 siRNA group 1 and(46.3±2.5)in CPLX2 siRNA group 2, which was lower than that in the blank group(88.0±2.0)and siRNA control group(77.0±4.4). Transwell invasion showed the number of penetrating cells was(29.7±2.5)in CPLX2 siRNA group 1 and(41.0±2.6)in CPLX2 siRNA group 2, which was lower than that in the blank group(74.7±3.1)and siRNA control group(68.7±1.5), and the difference was statistically significant(P<0.001). Conclusion CPLX2 is highly expressed in HCC tissues and knockdown of CPLX2 can inhibit the proliferation and migration of Huh7 cells, indicating that CPLX2 may play an important role in HCC tumorigenesis and progression.

Key words: Hepatocellular carcinoma, Complexin 2, Proliferation, Migration

中图分类号:

R735.7

孙薏丰,高玉,梁永媛,高杨. CPLX2在30例肝癌组织的表达及其对体外细胞增殖与侵袭的影响[J]. 山东大学学报 (医学版), 2020, 1(9): 34-39.

SUN Yifeng, GAO Yu, LIANG Yongyuan, GAO Yang. Expression of CPLX2 and its in vitro effects on the proliferation migration and invasion of hepatocellular carcinoma cells[J]. Journal of Shandong University (Health Sciences), 2020, 1(9): 34-39.

参考文献 25

[1]	Tsuru E, Oryu K, Sawada K, et al. Complexin 2 regulates secretion of immunoglobulin in antibody-secreting cells [J]. ImmunInflamm Dis, 2019, 7(4): 318-325.
[2]	Courtney NA, Bao H, Briguglio JS, et al. Synaptotagmin 1 clamps synaptic vesicle fusion in mammalian neurons independent of complexin [J]. Nature Communications, 2019, 10(1): 4076.
[3]	Kurokawa A, Narukawa M, Ohmoto M, et al. Expression of the synaptic exocytosis-regulating molecule complexin 2 in taste buds and its participation in peripheral taste transduction [J]. J Neurochem, 2015, 133(6): 806-814.
[4]	Zakharyan R, Atshemyan S, Boyajyan A. Risk and protective effects of the complexin-2 gene and gene-environment interactions in schizophrenia [J]. Recent Adv DNA Gene Seq, 2014, 8(1): 30-34.
[5]	Hass J, Walton E, Kirsten H, et al. Complexin2 modulates working memory-related neural activity in patients with schizophrenia [J]. Eur Arch Psychiatry Clin Neurosci, 2015, 265(2): 137-145.
[6]	Sia D, Villanueva A, Friedman SL, et al. Liver cancer cell of origin, molecular class, and effects on patient prognosis [J]. Gastroenterology, 2017, 152(4): 745-761.
[7]	Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries [J]. CA Cancer J Clin, 2018, 68(6): 394-424.
[8]	邵倩倩, 王景浦, 王庆杰. 抗凋亡转录因子在肝细胞肝癌中的表达及预后作用[J]. 山东大学学报(医学版), 2018, 56(12): 7-12. SHAO Qianqian, WANG Jingpu, WANG Qingjie. Expression and prognostic significance of apoptosis antagonizing transcription factor in hepatocellular carcinoma [J]. Journal of Shandong University(Health Sciences), 2018, 56(12): 7-12.
[9]	Chen Y, Mo L, Wang X, et al. TPGS-1000 exhibits potent anticancer activity for hepatocellular carcinoma in vitro and in vivo [J]. Aging(Albany NY), 2020, 12(2): 1624-1642.
[10]	Shen S, Peng H, Wang Y, et al. Screening for immune-potentiating antigens from hepatocellular carcinoma patients after radiofrequency ablation by serum proteomic analysis [J]. BMC Cancer, 2018, 18(1): 117.
[11]	Monsour Jr HP, Asham E, McFadden RS, et al. Hepatocellular carcinoma: the rising tide from east to west—a review of epidemiology, screening and tumor markers [J]. Translational Cancer Research, 2013, 2(6): 492-506.
[12]	Reghupaty SC, Sarkar D. Current status of gene therapy in hepatocellular carcinoma [J]. Review of. Cancers(Basel), 2019, 11(9). pii: E1265. doi: 10.3390/cancers11091265.
[13]	TsuruE, Oryu K, Sawada K, et al. Complexin 2 regulates secretion of immunoglobulin in antibody: ecreting cells [J]. Immunity, Inflammation and Disease, 2019, 7(4): 318-325.
[14]	López-Murcia FJ, Reim K, Jahn O, et al. Acute Complexin Knockout Abates Spontaneous and Evoked Transmitter Release [J]. Cell Rep, 2019, 26(10): 2521-2530.
[15]	Scholz N, Ehmann N, Sachidanandan D, et al. Complexin cooperates with Bruchpilot to tether synaptic vesicles to the active zone cytomatrix [J]. J Cell Biol, 2019, 218(3): 1011-1026.
[16]	Snead D, Eliezer D. Spectroscopic Characterization of Structure-Function Relationships in the Intrinsically Disordered Protein Complexin [J]. Methods Enzymol, 2018, 611: 227-286.
[17]	Ramos-Miguel A, Jones AA, Sawada K, et al. Frontotemporal dysregulation of the SNARE protein interactome is associated with faster cognitive decline in old age [J]. Neurobiol Dis, 2018, 114: 31-44.
[18]	Ramos-Miguel A, Beasley CL, Dwork AJ, et al. Increased SNARE protein-protein interactions in orbitofrontal and anterior cingulate cortices in schizophrenia [J]. Biol Psychiatry, 2015, 78(6): 361-373.
[19]	Begemann M, Grube S, Papiol S, et al. Modification of cognitive performance in schizophrenia by complexin 2 gene polymorphisms [J]. Arch Gen Psychiatry, 2010, 67(9): 879-888.
[20]	Glynn D, Gibson HE, Harte MK, et al. Clorgyline-mediated reversal of neurological deficits in a Complexin 2 knockout mouse [J]. Hum Mol Genet, 2010, 19(17): 3402-3412.
[21]	Radyushkin K, El-Kordi A, Boretius S, et al. Complexin2 null mutation requires a 'second hit' for induction of phenotypic changes relevant to schizophrenia [J]. Genes Brain Behav, 2010, 9(6): 592-602.
[22]	Sukumaran SK, Lewandowski BC, Qin Y, et al. Whole transcriptome profiling of taste bud cells [J]. Sci Rep, 2017, 7(1): 7595. doi: 10.1038/s41598-017-07746-z.
[23]	ó'Léime CS, Hoban AE, Hueston CM, et al. The orphan nuclear receptor TLX regulates hippocampal transcriptome changes induced by IL-1β[J]. Brain BehavImmun, 2018, 70: 268-279. doi: 10.1016/j.bbi.2018.03.006.
[24]	Makuuchi R, Terashima M, Kusuhara M, et al. Comprehensive analysis of gene mutation and expression profiles in neuroendocrine carcinomas of the stomach [J]. Biomed Res, 2017, 38(1): 19-27.
[25]	Komatsu H, Kakehashi A, Nishiyama N, et al. Complexin-2(CPLX2)as a potential prognostic biomarker in human lung high grade neuroendocrine tumors[J]. Cancer Biomark, 2013, 13(3): 171-180.

相关文章 15

[1]	王晓磊张海涛张辉郭成浩. 舒血宁注射液对高碘致培养血管内皮细胞损伤的保护作用[J]. 山东大学学报(医学版), 2209, 47(6): 38-.
[2]	殷悦,莫振飞,吴培昕,刘金霞,魏元辉,任佳博,李春笋. GPX1基因在肺癌中的表达特征及其对肺腺癌细胞增殖、迁移、侵袭、凋亡的影响[J]. 山东大学学报 (医学版), 2026, 64(1): 65-73.
[3]	韩觉明,王晖,吴倩,郑慧玲,朱琳. B4GALNT4促进肺腺癌细胞增殖、迁移和侵袭能力[J]. 山东大学学报 (医学版), 2025, 63(7): 23-31.
[4]	李观强,施昱诚,朱可涵,胡波,黄献琛,孙元,李笃信,张喜成. 蜗牛粘液来源的活性肽SK-14促进成纤维细胞的增殖和迁移[J]. 山东大学学报 (医学版), 2025, 63(11): 1-7.
[5]	刘振昆,吕纪玲,徐伟伟,马力天,张才擎. BALF tNGS检测及培养对NSCLC合并IPFD的诊断价值[J]. 山东大学学报 (医学版), 2025, 63(11): 36-45.
[6]	张洁,张芳芳,王靖楠,李泽宇,宋颖,李娜. circ_0000144在乳腺癌中的表达及其对乳腺癌细胞增殖、迁移和侵袭能力的影响[J]. 山东大学学报 (医学版), 2025, 63(1): 35-42.
[7]	张学宇,张学海,孙文青,刘晗,姜金波,刘寒,李远,陈晓梅. 重症新型冠状病毒肺炎伴侵袭性肺曲霉病及反复致命性消化道出血1例[J]. 山东大学学报 (医学版), 2024, 62(7): 56-61.
[8]	田丽君,桑玉洁,孙瑜婧,韩冰,秦成勇,祁建妮. 全身免疫炎症指数对原发性肝癌患者免疫检查点抑制剂治疗相关不良反应的预测价值[J]. 山东大学学报 (医学版), 2024, 62(6): 48-53.
[9]	曹华琳,贾彦召,曲莉,尹昕. CircFAT1调节miR-296-3p/MAPRE1轴对鼻咽癌细胞增殖、凋亡和放疗敏感性的影响[J]. 山东大学学报 (医学版), 2023, 61(9): 38-46.
[10]	金珊,高杰,谢玉姣,展垚,杜甜甜,王传新. 甲基转移酶PRMT5稳定USP15促进乳腺癌发生发展的作用[J]. 山东大学学报 (医学版), 2023, 61(7): 1-11.
[11]	靳新娟,左立平,邓展昊,李安宁,于德新. MRI影像组学对135例肝癌耐药蛋白PFKFB3的预测价值[J]. 山东大学学报 (医学版), 2023, 61(6): 79-86.
[12]	何静,严如根,武志红,李长忠. 消癥抑癌方对卵巢癌SKOV3细胞增殖、迁移的影响[J]. 山东大学学报 (医学版), 2023, 61(5): 1-10.
[13]	董相君,李娟,孔雪,李培龙,赵文静,梁怡然,王丽丽,杜鲁涛,王传新. 环状RNA hsa_circ_0008591对乳腺癌细胞生物学行为的影响[J]. 山东大学学报 (医学版), 2023, 61(2): 78-87.
[14]	赵舸,邹存华,宋冬冬,赵淑萍. 丹参酮IIA对子宫内膜癌细胞增殖与凋亡的影响[J]. 山东大学学报 (医学版), 2022, 60(9): 53-58.
[15]	张振伟,李佳,陈克明. IGF2BP2/m6A/ITGA5信号轴调控肾透明细胞增殖和迁移[J]. 山东大学学报 (医学版), 2022, 60(9): 74-84.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed