沉默WNT4基因对稳定转染PAX2基因大鼠肾小管上皮细胞迁移和侵袭能力的影响

doi:10.13481/j.1671-587x.20180618

Abstract

Abstract: Objective: To investigate the effects of WNT4 gene silecing on the migration and invasion of renal tubular epithelial cells of the rats stably transfected with paired box gene 2(PAX2) gene,and to clarify whether the PAX2 gene regulates the biological activities of renal tubular epithelial cells through WNT4 gene or not.Methods: The renal tubular epithelial cells transfected with PAX2 gene in the logarithmic phase were divided into stably transfected group and control group. The cells in stably transfected group were divided into stably transfected control group(without WNT4 siRNA silencing) and silencing 72 h group(with WNT4 siRNA silencing for 72 h). The relative expression amounts of PAX2 and WNT4 proteins in renal tubular epithelial cells of the rats in stably transfected group and control group were detected by Western blotting method. The relative expression amounts of WNT4 mRNA in renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group were detected by Real time PCR method. The migration rates of renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group were detected by cell wound scratch assay. The number of transmembrane renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group was detected by Transwell experiment.Results: The Western blotting results showed that the relative expression amounts of PAX2 and WNT4 proteins in renal tubular epithelial cells of the rats in stably transfected group were higher than those in control group(P<0.05). The Real time PCR results showed that the relative expression amount of WNT4 mRNA in renal tubular epithelial cells of the rats in silencing 72 h group was lower than that in stably transfected control group(P<0.05).The cell wound scratch assay results showed that the migration rate of renal tubular epithelial cells in silencing 72 h group after 10 h was lower than that in stably transfected control group,but there was no significant difference (P>0.05); the migration rate of renal tubular epithelial cells in silencing 72 h group after 18 h was lower than that in stably transfected control group(P<0.05).The Transwell experiment results showed that the number of transmembrane renal tubular epithelial cells in silencing 72 h group was lower than that in stably transfected control group(P<0.05).Conclusion: PAX2 gene can regulate the biological activities of renal tubular epithelial cells through WNT4 gene.

Key words: parired box gene 2, WNT4 gene, renal tubular epithelial cells, cell migration, cell invasion, transfection

CLC Number:

LI Li, GONG Liang, WU Yu bin. Effects of WNT4 gene silencing on migration and invasion of renal tubular epithelial cells of rats stably transfected with PAX2 gene[J].Journal of Jilin University(Medicine Edition), 2018, 44(06): 1212-1217.

References

[1] Kahraman K, Kiremitci S, Taskin S, et al. Expression pattern of PAX2 in hyperplastic and malignant endometrium[J]. Arch Gynecol Obstet, 2012, 286(1):173-178.
[2] El-Nahas AM. Plasticity of kidney cells:Role in kidney remodeling and scarring[J]. Kindey Int, 2003, 64(5):1553-1563.
[3] Narlis M, Grote D, Gaitan Y, et al.Pax2 and pax8 regulate branching morphogenesis and nephron differentiation in the developing kidney[J]. J Am Soc Nephrol, 2007, 18(4):1121-1129.
[4] Torban E, Goodyer P. What PAX genes do in the kidney[J]. Exp Nephrol, 1998,6(1):7-11.
[5] Torban E, Dziarmaga A, Iglesias D, et al. PAX2 activates WNT4 expression during mammalian kidney development[J]. J Biol Chem, 2006, 281(18):12705-12712.
[6] 李里,南晓娟,吴玉斌. WNT4基因沉默对PAX2诱导肾小管上皮细胞间充质转分化的影响[J]. 吉林大学学报:医学版,2013,39(4):720-725.
[7] 李里,宫亮,吴玉斌. PAX2稳转细胞系构建及对细胞迁移和侵袭能力的作用研究[J]. 中国全科医学, 2015,18(35):4325-4329.
[8] Lyons JP, Mueller UW, Ji H, et al. Wnt-4 activates the canonical beta-catenin-mediated Wnt pathway and binds Frizzled-6 CRD:functional implications of Wnt/beta-catenin activity in kidney epithelial cells[J]. Exp Cell Res, 2004, 298(2):369-387.
[9] Liu Y. New insights into epithelial-mesenchymal transition in kidney fibrosis[J].J Am Soc Nephrol, 2010, 21(2):212-222.
[10] Chilosi M, Poletti V, Zamo A, et al. Aberrant Wnt/beta-catenin pathway activation in idiopathic pulmonary fibrosis[J]. Am J Pathol, 2003, 162(5):1495-1502.
[11] Jiang F, Parsons CJ, Stefanovic B. Gene expression profile of quiescent and activated rat hepatic stellate cells implicates Wnt signaling pathway in activation[J]. J Hepatol, 2006, 45(3):401-409.
[12] Patel D, Shimomura A, Majumdar S, et al. The histone demethylase LSD1 regulates inner ear progenitor differentiation through interactions with Pax2 and the NuRD repressor complex[J]. PLoS One,2018,13(1):e0191689.
[13] Ma X, Ma Z, Jiao X, et al. Functional non-coding polymorphism in an EPHA2 promoter PAX2 binding site modifies expression and alters the MAPK and AKT pathways[J]. Sci Rep,2017,7(1):9992.
[14] Nguyen HT, Thomson AA, Kogan BA, et al. Expression of the Wnt gene family during late nephrogenesis and complete ureteral obstruction[J]. Lab Invest, 1999, 79(6):647-658.
[15] He W, Dai C, Li Y, et al. Wnt/beta-catenin signaling promotes renal interstitial fibrosis[J]. J Am Soc Nephrol, 2009, 20(4):765-776.
[16] Surendran K, McCaul SP, Simon TC. A role for Wnt-4 in renal fibrosis[J].Am J Physiol Renal Physiol, 2002, 282(3):F431-F441.
[17] Surendran K, Schiavi S, Hruska KA. Wnt-dependent beta-catenin signaling is activated after unilateral ureteral obstruction, and recombinant secreted frizzled-related protein 4 alters the progression of renal fibrosis[J]. J Am Soc Nephrol, 2005, 16(8):2373-2384.
[18] Chen Y, Zhang P, Tang P, et al. Wnt4 overexpression promotes thymoma development through a JNK-mediated planar cell polarity-like pathway[J]. Oncol Lett,2018,15(1):83-90.
[19] Wang L, Qing L, Liu H, et al. Mesenchymal stromal cells ameliorate oxidative stress-induced islet endothelium apoptosis and functional impairment via Wnt4-β-catenin signaling[J]. Stem Cell Res Ther,2017,8(1):188-122.
[20] 雷蕾,李良平,张虎.血清胱抑素C联合尿中性粒细胞明胶酶相关脂质运载蛋白对肝硬化腹水继发急性肾损伤的诊断价值[J].临床肝胆病杂志,2018,34(1):101-105.

Effects of WNT4 gene silencing on migration and invasion of renal tubular epithelial cells of rats stably transfected with PAX2 gene

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Metrics

Comments

Recommended 0

[1]	YUAN Huqin, LI Qiang. Inhibitory effects of silymarin on proliferation, migration and invasion of human gastric cancer SGC 7901 cells and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 988-993.
[2]	ZHAO Liyan, SONG Yang, CHEN Yong, LYU Xiaoyan, JIA Mingbo, LI Yunqian. Inhibitory effect of resveratrol on epithelial to mesenchymal transition of glioma U87 cells [J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 943-948.
[3]	YAN Hui, MA Shufei, DUAN Minghua, YAN Yuli, PAN Xin, LI Haiqing, ZHOU Changlong, ZHAO Tianyi. Influence of genipin in abilities of proliferation and invasion and apoptosis of gastric cancer SGC 7901 cells and their mechanisms in vitro [J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 924-928.
[4]	XING Shugang, ZHANG Lihong, JIN Minghua, LI Baihui, WU Zexin, FANG Bo, LI Wei. Effects of myricetin on proliferation and apoptosis of glioma GL261 cells in mice [J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 955-961.
[5]	ZHU Xiaohui, MENG Qin, FENG Shiyan. Expression of TP53 in villus tissue of unexplained recurrent spontaneous abortion and its effect on growth of human trophoblast cells [J]. Journal of Jilin University Medicine Edition, 2018, 44(04): 796-800.
[6]	YANG Dongye, JIAO Yang, TAN Bibo, DU Zhijian, HU Xiaojie, LIU Fei. Expression of Gli1 in gastric carcinoma tissue and its effect on biological characteristics of gastric carcinoma cells [J]. Journal of Jilin University Medicine Edition, 2018, 44(01): 63-67.
[7]	ZHAO Yanying, WANG Xiuyan, QIN Guotao, SUN Yuanjie, LIU Zhizhong, LI Ranwei. Effect of VEGF165b on biological characteristics of human hepatocellular carcinoma HepG₂ cells [J]. Journal of Jilin University Medicine Edition, 2018, 44(01): 101-105.
[8]	BI Ran, DU Yujun, WEI Wei, WANG Chunxi. Effects of andrographolide in proliferation,migration and apoptosis of renal cell carcinoma 786-0 cells [J]. Journal of Jilin University Medicine Edition, 2018, 44(01): 18-23.
[9]	LI Haobo, DU Yong, CHEN Yong, LI Wenjing. Influence of Notch 1 in expression of EGFR in tongue squamous cell carcinoma Tca8113 cells [J]. Journal of Jilin University Medicine Edition, 2017, 43(04): 715-719.
[10]	WANG Liping, FENG Ruo, ZOU Liyang, DING Yi. Influence of gallic acid in migration ability of human gastric carcinoma SGC-7901 cells [J]. Journal of Jilin University Medicine Edition, 2017, 43(04): 739-742.
[11]	CHEN Zhihua, LIN Suyong, HAN Hongjing, SU Xiaobao, CHEN Shaoqin, DAI Qibao. Effects of KISS1 gene transfected by lentivirus on proliferation, invasion, and migration abilities of human colorectal cancer HCT116 cells [J]. Journal of Jilin University Medicine Edition, 2017, 43(03): 577-581.
[12]	YU Haifan, GUO Chuanhui, LI Dangdang, ZHANG Hongliang, GENG Shuang, YANG Zhanqing, GUO Bin, YUE Zhanpeng. Effect of pleiotrophin on decidualization of uterine stromal cells in mice [J]. Journal of Jilin University Medicine Edition, 2016, 42(05): 855-859.
[13]	LI Fenge, XIANG Guoyan, KONG Fanli, HAO Feng, YANG Ziyan, ZHENG Yan, FANG Fang. Optimization of liposome-mediated transfection of pEGFP-N1 and its applification [J]. Journal of Jilin University Medicine Edition, 2016, 42(03): 612-616.
[14]	HAN Chunyao, ZHANG Bin, MA Xue, LIU Mingyuan, XUE Zhongyuan, HAO Lijing, GE Shuqing. Establishment of tongue squamous cell carcinoma TSCCA cell model with silencing α-catulin gene [J]. Journal of Jilin University Medicine Edition, 2016, 42(03): 481-485.
[15]	PAN Tao, GUO Caixia, JIN Minghua, LIU Xiaomei, LIU Ying, DU Haiying, SUN Zhiwei. Effects of SiO₂ nanoparticles on cell adhesion and migration of HL-7702 cells [J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 732-736.