[1]常海平,宋淑芳,郑健.过表达TPX2对人宫颈癌HeLa细胞侵袭和凋亡的影响[J].第三军医大学学报,2018,40(03):216-221.
 CHANG Haiping,SONG Shufang,ZHENG.Effect of overexpression of TPX2 gene on apoptosis and invasion in human cervical cancer HeLa cells[J].J Third Mil Med Univ,2018,40(03):216-221.
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过表达TPX2对人宫颈癌HeLa细胞侵袭和凋亡的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第03期
页码:
216-221
栏目:
基础医学
出版日期:
2018-02-15

文章信息/Info

Title:
Effect of overexpression of TPX2 gene on apoptosis and invasion in human cervical cancer HeLa cells
作者:
常海平宋淑芳郑健
内蒙古医科大学附属医院妇产科
Author(s):
CHANG Haiping SONG Shufang ZHENG

JianDepartment of Obstetrics and Gynecology, the Affiliated Hospital of Inner Mongolia Medical College, Hohhot, Inner Mongolia Autonomous Region, 010050, China

关键词:
HeLa细胞TPX2过表达细胞凋亡宫颈癌细胞侵袭
Keywords:
HeLa cells targeting protein for Xklp2 overexpression cervical cancer apoptosis invasion
分类号:
R394.3; R730.23; R737.33
文献标志码:
A
摘要:

目的     通过TPX2过表达慢病毒载体在人宫颈癌HeLa细胞过表达TPX2基因,在体外研究TPX2对人宫颈癌HeLa细胞侵袭和凋亡的作用及其相关机制。方法     构建TPX2过表达慢病毒载体(LV11-TPX2)及阴性对照(LV11NC),选取稳定感染过表达慢病毒载体(LV11TPX2)的HeLa细胞作为过表达组,将稳定感染(LV11-NC)的HeLa细胞作为阴性对照组,未感染病毒的人宫颈癌HeLa细胞作为空白对照组(CON)。采用Transwell基底膜侵袭实验测定各组细胞的侵袭能力,流式细胞术检测各组细胞的凋亡情况,Western blot检测细胞凋亡及侵袭相关蛋白质Bcl-2、Bax、Caspase-3、MMP-9、基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)及nm23-H1的表达。结果     成功构建TPX2过表达慢病毒载体(LV11-TPX2),可有效过表达TPX2 mRNA及蛋白质。与对照组比较, TPX2过表达组的HeLa细胞凋亡率明显减少(P<0.05),穿过Transwell小室基底膜细胞明显增加(P<0.01)。LV11-TPX2感染组HeLa细胞中凋亡相关蛋白Bcl-2的表达明显上调(P<0.05), Caspase-3及Bax的表达明显下调(P<0.05);侵袭相关蛋白质nm23-H1及TIMP-1水平明显下调(P<0.05),MMP9水平明显上调(P<0.05)。结论     在宫颈癌细胞过表达TPX2基因后,能抑制细胞凋亡,增强其侵袭能力,可能的机制为在宫颈癌细胞过表达TPX2能诱导凋亡和侵袭相关蛋白Caspase-3、Bax、nm23-H1及TIMP-1水平下调,Bcl-2及MMP-9水平上调。

Abstract:

Objective     To investigate the effect of targeting protein for Xklp2 (TPX2) on the invasion and apoptosis of human cervical carcinoma HeLa cells by lentiviral vectors carrying TPX2 and to explore its possible mechanism. Methods    Lentiviral vectors carrying TPX2 (LV11-TPX2) and negative control were constructed respectively. HeLa cells stably infected with LV11-TPX2 served experimental group, the cells infected with LV11-NC as negative control group, and the cells without infection as the blank control group. Cell invasion and apoptosis were determined by Transwell chamber test and flow cytometry, respectively. The expression levels of Bcl-2, Bax, Caspase-3, MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) and nm23H1 were detected by Western blotting. Results    The lentiviral vector LV11-TPX2 was successfully constructed, and its infection into HeLa cells resulted in effectively overexpressed TPX2 at mRNA and protein levels. Compared with the control group, the apoptotic rate of LV11-TPX2 group was significantly reduced (P<0.05), and the cells through the Transwell chamber basement membrane was significantly increased (P<0.01). Overexpression of TPX2 gene could down-regulate the protein levels of Caspase-3, Bax, nm23-H1 and TIMP-1, and up-regulate the expression of Bcl-2 and MMP-9 in HeLa cells (P<0.05). Conclusion     Overexpression of TPX2 gene inhibits cell apoptosis and promotes invasive ability in cervical carcinoma cells, which may be through down-regulating the expression of Caspase-3, Bax, nm23-H1 and TIMP-1, and up-regulating the expression of Bcl-2 and MMP-9.

参考文献/References:

[1]PREZ DE CASTRO I, MALUMBRES M. Mitotic stress and chromosomal instability in cancer: the case for TPX2[J]. Genes Cancer, 2012, 3(11/12): 721-730. DOI:10.1177/1947601912473306.
[2]刘万伟, 李恩亮, 邬林泉. TPX2在细胞间期参与DNA损伤反应及其与肿瘤发生关系的研究进展[J]. 广东医学, 2016, 37(8): 1245-1248.
LIU W W, LI E L, WU L Q. TPX2 involved in the DNA damage response in the interphase of the cell and its relationship with tumor research progress [J]. Guangdong Med J, 2016, 37(8): 1245-1248.
[3]常海平,王敬芝,田原,等.TPX2在宫颈癌中的表达及意义[J].基础医学与临床,2012,32(5):561-565.
CHANG H P, WANG J Z, TIAN Y, et al. Expression of TPX2 in cervical carcinoma and its significance[J]. Basic Clin Med, 2012, 32(5): 561-565.
[4]常海平, 杨彩荣, 郑健. 人TPX2基因过表达慢病毒质粒的构建及其人宫颈癌细胞稳定表达株的筛选[J]. 中国生物制品学杂志, 2017, 30(6): 607-612.
CHANG H P, YANG C R, ZHENG J. Construction of lentivirus vector for overexpression of human TPX2 gene and screening of human cervical cancer cell line stably expressing TPX2[J]. Chin J Biol, 2017, 30(6): 607-612.
[5]常海平,杨彩荣,赵荣伟,等.慢病毒介导的TPX2沉默对人宫颈癌HeLa细胞凋亡和侵袭的影响及机制[J].中国细胞生物学学报,2017, 39(11): 1415-1421. DOI: 10.11844/cjcb.2017.11.0152.
CHANG H P, YANG C R,ZHAO R W, et al.The effect of silencing TPX2 gene expression mediated by lentiviral on apoptosis and invasion in human cervical cancer HeLa cells and its mechanism[J]. Chin J Cell Biology, 2017, 39(11): 1415-1421. DOI: 10.11844/cjcb.2017.11.0152.
[6]GRUSS O J, WITTMANN M, YOKOYAMA H, et al. Chromosomeinduced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells[J]. Nat Cell Biol, 2002, 4(11): 871-879. DOI:10.1038/ncb870.
[7]YAN L, LI S, XU C, et al. Target protein for Xklp2 (TPX2), a microtubulerelated protein, contributes to malignant phenotype in bladder carcinoma[J]. Tumour Biol, 2013, 34(6): 4089-4100. DOI:10.1007/s132770131000z.
[8]HSU P K, CHEN H Y, YEH Y C, et al. TPX2 expression is associated with cell proliferation and patient outcome in esophageal squamous cell carcinoma[J]. J Gastroenterol, 2014, 49(8): 1231-1240. DOI:10.1007/s0053501308706.
[9]HUANG C, HAN Z, WU D. Effects of TPX2 gene on radiotherapy sensitization in breast cancer stem cells[J]. Oncol Lett, 2017, 14(2): 1531-1535. DOI:10.3892/ol.2017.6277.
[10]SCHNEIDER M A, CHRISTOPOULOS P, MULEY T, et al. AURKA, DLGAP5, TPX2, KIF11 and CKAP5: five specific mitosisassociated genes correlate with poor prognosis for nonsmall cell lung cancer patients [J]. Int J Oncol, 2017, 50(2):365-372.DOI: 10.3892/ijo.2017.3834.
[11]PAN H W, SU H H, HSU C W, et al. Targeted TPX2 increases chromosome missegregation and suppresses tumor cell growth in human prostate cancer[J]. Onco Targets Ther, 2017,10:3531-3543. DOI: 10.2147/OTT.S136491.
[12]TOMII C, INOKUCHI M, TAKAGI Y, et al. TPX2 expression is associated with poor survival in gastric cancer[J]. World J Surg Oncol, 2017,15(1):14.DOI: 10.1186/s129570161095y.
[13]HSU C W, CHEN Y C, SU H H, et al. Targeting TPX2 suppresses the tumorigenesis of hepatocellular carcinoma cells resulting in arrested mitotic phase progression and increased genomic instability[J]. J Cancer, 2017, 8(8):1378-1394.DOI: 10.7150/jca.17478.
[14]黄维, 陈洁仪, 黄俊勇, 等. TRPM8对人脑胶质瘤细胞U251细胞凋亡和细胞增殖的影响[J]. 热带医学杂志, 2015, 15(2): 203-206.
HUANG W, CHEN J Y, HUANG J Y, et al. Effect of trpm8 on proliferation and apoptosis of human brain gliomacell u251[J]. J Trop Med, 2015, 15(2): 203-206.
[15]HUANG Q, LI F, LIU X, et al. Caspase 3mediated stimulation of tumor cell repopulation during cancer radiotherapy[J]. Nat Med, 2011, 17(7): 860-866. DOI:10.1038/nm.2385.
[16]李琴, 郭云良, 李震, 等. 胡黄连苷Ⅱ对大鼠脑缺血/再灌注损伤Caspase3和PARP表达的影响[J]. 中国药理学通报, 2010, 26(3): 342-345.
LI Q, GUO Y L, LI Z, et al. The interference of picroside Ⅱ on the expressions of caspase3 and parp following cerebral ischemia reperfusion injury in rats[J]. Chin Pharmacol Bull, 2010, 26(3): 342-345.
[17]董雅洁, 高维娟. bcl2、bax、caspase3在细胞凋亡中的作用及其关系[J].中国老年学杂志, 2012,32(11): 4828-4830.
DONG Y J, GAO W J. The roles of BCL2, bax and caspase3 in the apoptosis and their relationships[J]. Chin J Gerontol, 2012,32(11): 4828-4830.
[18]PYTLIAK M, VARGOV V, MECHROV V. Matrix metalloproteinases and their role in oncogenesis: a review[J]. Onkologie, 2012, 35(1/2): 49-53. DOI:10.1159/000336304.
[19]HARTSOUGH M T. Nm23H1 metastasis suppressor phosphorylation of kinase suppressor of ras via a histidine protein kinase pathway[J]. J Biol Chem,2002,277(35):32389-32399. DOI: 10.1074/jbc.M203115200.

更新日期/Last Update: 2018-02-08