沉默T-cadherin对ox-LDL诱导人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响

doi:10.11958/20220832

天津医药 ›› 2023, Vol. 51 ›› Issue (1): 8-13.doi: 10.11958/20220832

沉默T-cadherin对ox-LDL诱导人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响

王海娇¹(), 何红美¹, 祁麟¹, 崔玉娇¹, 肖春辉², 王毅¹^,^∆()

1 石家庄市第四医院检验科，河北省母胎医学重点实验室（邮编050032）
2 石家庄市第四医院产一科

收稿日期:2022-05-26 修回日期:2022-06-17 出版日期:2023-01-15 发布日期:2023-01-17
通讯作者: 王毅 E-mail:wanghaijiao527@163.com;13060509@qq.com
作者简介:王海娇（1981），女，主管检验师，主要从事产科疾病方面研究。E-mail：wanghaijiao527@163.com
基金资助:
河北省医学科学研究课题计划项目(20201380)

Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo

WANG Haijiao¹(), HE Hongmei¹, QI Lin¹, CUI Yujiao¹, XIAO Chunhui², WANG Yi¹^,^∆()

1 Department of Clinical Laboratory, The Fourth Hospital of Shijiazhuang, Hebei Provincial Key Laboratory of Maternal-Fetal Medicine
2 Department of First Obstetrics, The Fourth Hospital of Shijiazhuang, Shijiazhuang 050032, China

Received:2022-05-26 Revised:2022-06-17 Published:2023-01-15 Online:2023-01-17
Contact: WANG Yi E-mail:wanghaijiao527@163.com;13060509@qq.com

王海娇, 何红美, 祁麟, 崔玉娇, 肖春辉, 王毅. 沉默T-cadherin对ox-LDL诱导人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响[J]. 天津医药, 2023, 51(1): 8-13.

WANG Haijiao, HE Hongmei, QI Lin, CUI Yujiao, XIAO Chunhui, WANG Yi. Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo[J]. Tianjin Medical Journal, 2023, 51(1): 8-13.

摘要/Abstract

摘要：

目的探讨沉默T-钙黏蛋白（T-cadherin）对氧化低密度脂蛋白（ox-LDL）诱导的人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响。方法将人绒毛膜滋养层细胞HTR-8/SVneo分为空白对照组、ox-LDL组、T-cadherin小干扰RNA（siRNA）阴性对照组、T-cadherin siRNA组。各组细胞转染后，实时荧光定量PCR检测各组T-cadherin mRNA水平;MTT法检测HTR-8/SVneo细胞增殖情况;平板克隆形成实验检测细胞克隆形成情况;流式细胞仪检测细胞凋亡情况;Transwell小室实验、划痕实验分别检测细胞侵袭、迁移能力;蛋白免疫印迹实验检测细胞胱天蛋白酶-3（Caspase-3）、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）和基质金属蛋白酶-2（MMP-2）、MMP-9蛋白表达水平。结果与空白对照组相比，ox-LDL组和T-cadherin siRNA阴性对照组T-cadherin mRNA、细胞增殖抑制率、凋亡率、Caspase-3、Bax蛋白表达水平升高，克隆形成率、侵袭细胞数、划痕愈合率、Bcl-2、MMP-2、MMP-9蛋白表达水平降低（P<0.05）;与ox-LDL组和T-cadherin siRNA阴性对照组相比，T-cadherin siRNA组T-cadherin mRNA、细胞增殖抑制率、凋亡率、Caspase-3、Bax蛋白表达水平降低，克隆形成率、侵袭细胞数、划痕愈合率、Bcl-2、MMP-2、MMP-9蛋白表达水平升高（P<0.05）。结论沉默T-cadherin可抑制ox-LDL诱导的HTR-8/SVneo细胞异常凋亡，并促进细胞的增殖、侵袭、迁移能力。

关键词: 钙黏着糖蛋白类, 脂蛋白类，IDL, 滋养层, 细胞增殖, 细胞凋亡, T-钙黏蛋白, 氧化低密度脂蛋白

Abstract:

Objective To investigate the effect of silencing T-cadherin on oxidized low density lipoprotein (ox-LDL) induced biological behavior of human chorionic trophoblast HTR-8/SVneo cells. Methods Human chorionic trophoblast cells HTR-8/SVneo were divided into the blank control group, the ox-LDL group, the T-cadherin small interfering RNA (siRNA) negative control group and the T-cadherin siRNA group. After each group was treated accordingly, the expression levels of T-cadherin messenger RNA (mRNA) in HTR-8/SVneo cells were detected by real-time fluorescence quantitative PCR in the four groups. The proliferation of cells in each group was detected by tetramethylazolate method. The clone formation of each group was detected by plate clone formation experiment. The apoptosis of each group was detected by flow cytometry. Transwell chamber test and scratch test were used to detect the invasion and migration ability of cells. The protein expression levels of Caspase-3, Bcl-2 related X protein (Bax), B cell lymphoma-2 (Bcl-2) and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) of cells in each group were detected by Western blot assay. Results Compared with the blank control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were increased significantly in the ox-LDL group and the T-cadherin siRNA negative control group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were decreased significantly (P<0.05). Compared with the ox-LDL group and the T-cadherin siRNA negative control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were decreased significantly in the T-cadherin siRNA group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were increased significantly (P<0.05). Conclusion Silencing T-cadherin can inhibit ox-LDL induced abnormal apoptosis of HTR-8/SVneo cells, and promote cell proliferation, invasion and migration.

Key words: cadherins, lipoproteins, IDL, trophoblasts, cell proliferation, apoptosis, T-cadherin, oxidized low density lipoprotein

中图分类号:

R714.43

图/表 8

表1 各组HTR-8/SVneo细胞增殖抑制率、克隆形成率比较（n=6，%，$\bar{x}±s$）

Tab.1 Comparison of proliferation inhibition rate and clone formation rate between the four groups of HTR-8/SVneo cells

组别	细胞增殖抑制率	细胞克隆形成率
空白对照组	10.96±1.68	39.26±4.35
ox-LDL组	32.64±4.11^a	15.42±2.37^a
T-cadherin siRNA阴性对照组	31.23±3.97^a	16.85±2.16^a
T-cadherin siRNA组	20.60±3.06^bc	28.08±3.18^bc
F	55.031^＊＊	75.487^＊＊

图1 各组HTR-8/SVneo细胞克隆形成形态学观察（结晶紫染色，×40） A：空白对照组;B：ox-LDL组;C：T-cadherin siRNA阴性对照组;D：T-cadherin siRNA组。

Fig.1 Morphological observation of HTR-8/SVneo cell clone formation in each group (crystal violet staining, ×40）

图2 各组HTR-8/SVneo细胞凋亡情况 A：空白对照组;B：ox-LDL组;C：T-cadherin siRNA阴性对照组;D：T-cadherin siRNA组。

Fig.2 Apoptosis of HTR-8/SVneo cells in each group

表2 各组HTR-8/SVneo细胞侵袭细胞数和划痕愈合率比较（n=6，$\bar{x}±s$）

Tab.2 Comparison of the number of invasive cells and scratch healing rate of HTR-8/SVneo cells between the four groups

组别	侵袭细胞数（个/视野）	划痕愈合率（%）
空白对照组	140.22±22.95	25.66±4.30
ox-LDL组	65.47±8.04^a	9.08±1.09^a
T-cadherin siRNA阴性对照组	66.06±8.06^a	9.12±1.13^a
T-cadherin siRNA组	98.53±10.03^bc	20.11±3.77^bc
F	39.563^＊＊	46.731^＊＊

图3 各组HTR-8/SVneo细胞侵袭情况（结晶紫染色，×200） A：空白对照组;B：ox-LDL组;C：T-cadherin siRNA阴性对照组;D：T-cadherin siRNA组。

Fig.3 Invasion of HTR-8/SVneo cells in each group (crystal violet staining, ×200)

图4 各组HTR-8/SVneo细胞迁移情况（×100） A：空白对照组;B：ox-LDL组;C：T-cadherin siRNA阴性对照组;D：T-cadherin siRNA组。

Fig.4 Migration of HTR-8/SVneo cells in each group (×100)

图5 各组HTR-8/SVneo细胞Caspase-3、Bax、Bcl-2、MMP-2、MMP-9蛋白印迹图 A：空白对照组;B：ox-LDL组;C：T-cadherin siRNA阴性对照组;D：T-cadherin siRNA组。

Fig.5 Western blot results of Caspase-3, Bax, Bcl-2, MMP-2 and MMP-9 protein of HTR-8/SVneo cells in each group

表3 各组HTR-8/SVneo细胞Caspase-3、Bax、Bcl-2、MMP-2、MMP-9蛋白表达水平比较（n=6，$\bar{x}±s$）

Tab.3 Comparison of expression levels of Caspase-3, Bax, Bcl-2, MMP-2 and MMP-9 protein in HTR-8/SVneo cells between the four groups

组别	Caspase-3	Bax	Bcl-2	MMP-2	MMP-9
空白对照组	0.96±0.09	1.01±0.10	1.03±0.09	1.02±0.11	0.97±0.12
ox-LDL组	1.65±0.18^a	1.70±0.18^a	0.21±0.02^a	0.26±0.04^a	0.31±0.04^a
T-cadherin siRNA阴性对照组	1.62±0.17^a	1.72±0.19^a	0.18±0.03^a	0.28±0.04^a	0.29±0.03^a
T-cadherin siRNA组	1.17±0.14^bc	1.35±0.13^bc	0.66±0.07^bc	0.73±0.08^bc	0.72±0.07^bc
F	31.227^＊＊	28.419^＊＊	274.909^＊＊	150.516^＊＊	120.541^＊＊

参考文献 23

[1]	ZHAO X, LIU F, ZHANG J, et al. LINC01128-miR-16 interaction regulates the migration and invasion of human chorionic trophoblast cells[J]. Hypertens Pregnancy, 2021, 40(2):152-161. doi:10.1080/10641955.2021.1917602.
[2]	LIU G, JIA J, ZHONG J, et al. TCDD-induced IL-24 secretion in human chorionic stromal cells inhibits placental trophoblast cell migration and invasion[J]. Reprod Toxicol, 2022, 108:10-17. doi:10.1016/j.reprotox.2022.01.001.
[3]	ZHANG J, LIU X, GAO Y. Abnormal H3K27 histone methylation of RASA1 gene leads to unexplained recurrent spontaneous abortion by regulating Ras-MAPK pathway in trophoblast cells[J]. Mol Biol Rep, 2021, 48(6):5109-5119. doi:10.1007/s11033-021-06507-6.
[4]	王海臻, 蔡丹纯, 廖丹丹, 等. 内质网应激介导滋养细胞凋亡在妊娠期肝内胆汁淤积症中的作用及机制[J]. 南方医科大学学报, 2018, 38(5):572-577.
	WANG H Z, CAI D C, LIAO D D, et al. Role of endoplasmic reticulum stress-induced apoptosis of trophoblasts in intrahepatic cholestasis during pregnancy[J]. J South Med Univ, 2018, 38(5):572-577. doi:10.3969/j.issn.1673-4254.2018.05.011.
[5]	SHAN L, HOU X. Circular RNA hsa_circ_0026552 inhibits the proliferation, migration and invasion of trophoblast cells via the miR-331-3p/TGF-βR1 axis in pre-eclampsia[J]. Mol Med Rep, 2021, 24(5):798. doi:10.3892/mmr.2021.12438.
[6]	DI PALO A, SINISCALCHI C, POLITO R, et al. microRNA-377-3p downregulates the oncosuppressor T-cadherin in colorectal adenocarcinoma cells[J]. Cell Biol Int, 2021, 45(8):1797-1803. doi:10.1002/cbin.11605.
[7]	LIN J, CHEN Z, HUANG Z, et al. Upregulation of T-cadherin suppresses cell proliferation, migration and invasion of gastric cancer in vitro[J]. Exp Ther Med, 2017, 14(5):4194-4200. doi:10.3892/etm.2017.5090.
[8]	ZHAO J, YANG T, JI J, et al. Garcinol exerts anti-cancer effect in human cervical cancer cells through upregulation of T-cadherin[J]. Biomed Pharmacother, 2018, 107:957-966. doi:10.1016/j.biopha.2018.08.060.
[9]	李夏芳. 子痫前期胎盘组织中Wnt/β-catenin信号通路相关分子的含量及其与滋养细胞凋亡的相关性[J]. 海南医学院学报, 2017, 23(17):2388-2391.
	LI X F. The contents of Wnt/β-catenin signal ing pathway-related molecules in the preeclampsia pla-centa tissue and the correlation with the trophocyte apoptosis[J]. Journal of Hainan Medical University, 2017, 23(17):2388-2391. doi:10.13210/j.cnki.jhmu.20170829.008.
[10]	CARVAJAL L, CANTIN C, FUENZALIDA B, et al. Preeclampsia and ox-LDL modify the expression of autophagy markers in placenta and first trimester trophoblast cell line impairing trophoblast invasion and migration[J]. Placenta, 2019, 83(1):104-105. doi:10.1016/j.placenta.2019.06.331.
[11]	张妍. 自噬对氧化低密度脂蛋白诱导滋养细胞炎性反应的保护作用及其与子痫前期发病的关系[D]. 青岛: 青岛大学, 2017.
	ZHANG Y. Protective effect of autophagy on oxidative low density lipoprotein induced trophoblast inflammatory response and its relationship with preeclampsia[D]. Qingdao: Qingdao University, 2017.
[12]	ZHANG X, WANG Z, LI W, et al. MicroRNA-217-5p ameliorates endothelial cell apoptosis induced by ox-LDL by targeting CLIC4[J]. Nutr Metab Cardiovasc Dis, 2020, 30(3):523-533. doi:10.1016/j.numecd.2019.09.027.
[13]	刘琳, 柴志勇, 刁云辉, 等. 当归多糖对氧化型低密度脂蛋白诱导的血管内皮细胞损伤的保护作用研究[J]. 中国临床药理学杂志, 2020, 36(7):818-821.
	LIU L, CHAI Z Y, DIAO Y H, et al. Protective effect of angelica polysaccharide on vascular endothelial cell injury induced by oxidized low density lipoprotein[J]. Chin J Clin Pharmacol, 2020, 36(7):818-821. doi:10.13699/j.cnki.1001-6821.2020.07.026.
[14]	LOMBARDELLI L, LOGIODICE F, KULLOLLI O, et al. At embryo implantation site IL-35 secreted by trophoblast,polarizing T cells towards IL-35⁺ IL-10⁺ IL-4⁺ Th2-type cells,could favour fetal allograft tolerance and pregnancy success[J]. Int J Mol Sci, 2022, 23(9):4926. doi:10.3390/ijms23094926.
[15]	SHI S, TAN Q, LIANG J, et al. Placental trophoblast cell-derived exosomal microRNA-1290 promotes the interaction between endometrium and embryo by targeting LHX6[J]. Mol Ther Nucleic Acids, 2021, 26:760-772. doi:10.1016/j.omtn.2021.09.009.
[16]	张悦, 冯颖, 马芳. 早期停育胚胎的滋养层细胞相关基因与特征分析[J]. 遗传, 2020, 42(10):1004-1016.
	ZHANG Y, FENG Y, MA F. Related genes and characteristic analysis of trophoblast cells during early embryo developmental cessation[J]. Hereditas, 2020, 42(10):1004-1016. doi:10.16288/j.yczz.20-144.
[17]	ALVES P, AMARAL C, TEIXEIRA N, et al. Cannabidiol disrupts apoptosis, autophagy and invasion processes of placental trophoblasts[J]. Arch Toxicol, 2021, 95(10):3393-3406. doi:10.1007/s00204-021-03122-z.
[18]	周小波, 石书明, 张华. 线粒体自噬受损引起的ROS清除障碍导致子痫前期患者滋养细胞过度凋亡[J]. 重庆医科大学学报, 2017, 42(3):315-318.
	ZHOU X B, SHI S M, ZHANG H. Impaired mitophagy is involved in the trophoblastic excessive apoptosis in preeclampsia caused by ROS[J]. Journal of Chongqing Medical University, 2017, 42(3):315-318. doi:10.13406/j.cnki.cyxb.001218.
[19]	BIYIK I, KALKAN U, SIMSEK S. The deep infiltrating endometriosis tissue has lower T-cadherin,E-cadherin,progesterone receptor and oestrogen receptor than endometrioma tissue[J]. Taiwan J Obstet Gynecol, 2021, 60(6):1059-1065. doi:10.1016/j.tjog.2021.09.017.
[20]	FUKUDA S, KITA S, MIYASHITA K, et al. Identification and clinical associations of 3 forms of circulating T-cadherin in human serum[J]. J Clin Endocrinol Metab, 2021, 106(5):1333-1344. doi:10.1210/clinem/dgab066.
[21]	陈杰, 王少敏, 王锋, 等. T-cadherin在卵巢癌中的表达及其对卵巢癌细胞生物学行为的影响[J]. 山西医科大学学报, 2021, 52(9):1115-1120.
	CHEN J, WANG S M, WANG F, et al. Expression of T-cadherin in ovarian cancer and its effect on biological behaviors of ovarian cancer cells[J]. Journal of Shanxi Medical University, 2021, 52(9):1115-1120. doi:10.13753/j.issn.1007-6611.2021.09.005.
[22]	AAMAZADEH F, OSTADRAHIMI A, RAHBAR SAADAT Y, et al. Bitter apricot ethanolic extract induces apoptosis through increasing expression of Bax/Bcl-2 ratio and caspase-3 in PANC-1 pancreatic cancer cells[J]. Mol Biol Rep, 2020, 47(3):1895-1904. doi:10.1007/s11033-020-05286-w.
[23]	CHEN T, CHEN Z, LIAN X, et al. MUC 15 promotes osteosarcoma cell proliferation,migration and invasion through Livin,MMP-2/MMP-9 and Wnt/β-catenin signal pathway[J]. J Cancer, 2021, 12(2):467-473. doi:10.7150/jca.49641.

沉默T-cadherin对ox-LDL诱导人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响

Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo

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