您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报 (医学版) ›› 2020, Vol. 58 ›› Issue (9): 34-39.doi: 10.6040/j.issn.1671-7554.0.2020.0748

• • 上一篇    下一篇

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

孙薏丰1,高玉2,梁永媛1,高杨1   

  1. 1. 山东大学齐鲁医院检验科, 山东 济南 250012;2.青岛市市北区妇幼保健计划生育服务中心, 山东 青岛 266032
  • 出版日期:2020-09-10 发布日期:2020-08-30
  • 通讯作者: 高杨. E-mail:gy2469@163.com

Expression of CPLX2 and its in vitro effects on the proliferation migration and invasion of hepatocellular carcinoma cells

SUN Yifeng1, GAO Yu2, LIANG Yongyuan1, GAO Yang1   

  1. 1. Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    2. Maternal and Child Health Care of Family Planning Service Center of Shibei District, Qingdao 266032, Shandong, China
  • Online:2020-09-10 Published:2020-08-30

PDF (PC)

176

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
[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.
[1] 李涵,付婷婷,张磊,延冰,孙涛,郭峰,尹晓. 过氧化物酶增殖物激活受体γ激动剂对24例肥胖症患者米色脂肪细胞分化的影响[J]. 山东大学学报 (医学版), 2020, 58(9): 8-13.
[2] 张宝文,雷香丽,李瑾娜,罗湘俊,邹容. miR-21-5p靶向调控TIMP3抑制2型糖尿病肾病小鼠肾脏系膜细胞增殖及细胞外基质堆积[J]. 山东大学学报 (医学版), 2020, 58(7): 7-14.
[3] 付洁琦,张曼,张晓璐,李卉,陈红. Toll样受体4抑制过氧化物酶体增殖物激活受体γ加重血脂蓄积的分子机制[J]. 山东大学学报 (医学版), 2020, 58(7): 24-31.
[4] 马青源,蒲沛东,韩飞,王超,朱洲均,王维山,史晨辉. miR-27b-3p调控SMAD1对骨肉瘤细胞增殖、迁移和侵袭作用的影响[J]. 山东大学学报 (医学版), 2020, 58(7): 32-37.
[5] 李宁,李娟,谢艳,李培龙,王允山,杜鲁涛,王传新. 长链非编码RNA AL109955.1在80例结直肠癌组织中的表达及对细胞增殖与迁移侵袭的影响[J]. 山东大学学报 (医学版), 2020, 58(7): 38-46.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发