[1]刘鑫,刘小兵,刘骞,等.钙激活核苷酸酶1敲低对肾透明细胞癌769-P细胞增殖和迁移的影响[J].第三军医大学学报,2018,40(06):473-478.
 LIU Xin,LIU Xiaobing,LIU Qian,et al.Silencing calcium-activated nucleotidase 1 inhibits proliferation and migration of clear cell renal cell carcinoma 769-P cells[J].J Third Mil Med Univ,2018,40(06):473-478.
点击复制

钙激活核苷酸酶1敲低对肾透明细胞癌769-P细胞增殖和迁移的影响(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第06期
页码:
473-478
栏目:
基础医学
出版日期:
2018-03-30

文章信息/Info

Title:
Silencing calcium-activated nucleotidase 1 inhibits proliferation and migration of clear cell renal cell carcinoma 769-P cells
作者:
刘鑫刘小兵刘骞王青青杨振兴李龙坤
陆军军医大学(第三军医大学)第二附属医院泌尿外科
Author(s):
LIU Xin LIU Xiaobing LIU Qian WANG Qingqing YANG Zhenxing LI Longkun

Department of Urology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China

关键词:
肾透明细胞癌钙激活核苷酸酶1细胞增殖细胞迁移细胞凋亡
Keywords:
clear cell renal cell carcinoma  calcium-activated nucleotidase 1 proliferationmigrationapoptosis
分类号:
R394.3;R730.23;R737.11
文献标志码:
A
摘要:

目的    探究钙激活核苷酸酶1(calciumactivated nucleotidase 1, CANT1)对人肾透明细胞癌769P细胞系增殖和迁移的影响,并对其作用机制进行初步探讨。方法    免疫组化检测临床肿瘤及癌旁组织CANT1的表达差异。包装携带干扰CANT1的shRNA及空白对照shRNA慢病毒,并转染769-P 细胞系,采用RT-PCR和Western blot分别在mRNA及蛋白水平验证shRNA干扰效率。稳定转染后,采用CCK-8和划痕实验分别检测细胞增殖、迁移能力,采用流式细胞仪检测细胞周期、细胞凋亡情况。结果    成功建立稳定转染的CANT1干扰细胞系。免疫组化显示CANT1在癌周正常肾组织中仅表达于远曲小管,近曲小管无或者仅有少量表达,而在肾透明细胞癌组织中,CANT1呈弥漫高表达状态。CANT1干扰组细胞较对照组细胞增殖明显减慢(P<0.01),迁移能力明显减弱(P<0.05),干扰组细胞周期阻滞于S期,细胞凋亡增多(P<0.05)。结论    CANT1敲低能明显抑制肾透明细胞癌769P细胞系的增殖和迁移,使细胞周期阻滞于S期,凋亡细胞明显增多。

Abstract:

Objective    To determine the effect of calcium-activated nucleotidase 1 (CANT1)on the proliferation and migration in human clear cell renal cell carcinoma 769-P cells, and preliminarily investigate its underlying mechanism. Methods    The expression of CANT1 was detected by immunohistochemistry in the intraoperatively obtained tumor tissues and paracancerous normal tissues. Lentiviral vectors carrying shRNA interfering CANT1 and blank control shRNA were used to infect the 769-P cells, then RT-PCR and Western blotting were adopted to verify the interference efficiency at mRNA and protein levels. After stable transfection, cell proliferation, migration,cell cycle distribution and apoptosis were measured. Results    The 769-P cells with stable CANT1 knockdown were successfully established.Immunohistochemistry showed that CANT1 was expressed only in the distant renal tubules of normal renal tissues, but was with little or no expression in the proximal renal tubule.In the tissues of clear cell renal cell carcinoma, the protein was intensively expressed. CANT1 knockdown significantly inhibited the cell proliferation (P<0.01) and cell migration by wound healing assay(P<0.01), arrested the cells at S phase, and enhanced cell apoptosis in the 769P cells after transfection with lentiviral vectors carrying shRNA CANT1 than those transfected with blank control shRNA(P<0.05). Conclusion    CANT1 knockdown can significantly inhibit the proliferation and migration, arrest cell cycle at S stage and promote the apoptosis in clear cell renal cell carcinoma 769-P cells.

参考文献/References:

[1]SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2017[J]. CA Cancer J Clin, 2017, 67(1): 7-30. DOI: 10.3322/caac.21387.
[2]MOTZER R J, BANDER N H, NANUS D M. Renalcell carcinoma[J]. N Engl J Med, 1996, 335(12): 865-875. DOI: 10.1056/NEJM199609193351207.
[3]JANZEN N K, KIM H L, FIGLIN R A, et al. Surveillance after radical or partial nephrectomy for localized renal cell carcinoma and management of recurrent disease[J]. Urol Clin North Am, 2003,30(4): 843-852.
[4]SMITH T M, HICKSBERGER C A, KIM S, et al. Cloning, expression, and characterization of a soluble calciumactivated nucleotidase, a human enzyme belonging to a new family of extracellular nucleotidases[J]. Arch Biochem Biophys, 2002, 406(1): 105-115.
[5]LACCONE F, SCHONER K, KRABICHLER B, et al. Desbuquois dysplasia type I and fetal hydrops due to novel mutations in the CANT1 gene[J]. Eur J Hum Genet, 2011, 19(11): 1133-1137. DOI: 10.1038/ejhg.2011.101.
[6]HUBER C, OULES B, BERTOLI M, et al. Identification of CANT1 mutations in Desbuquois dysplasia[J]. Am J Hum Genet, 2009, 85(5): 706-710. DOI: 10.1016/j.ajhg.2009.10.001.
[7]FURUICHI T, DAI J, CHO T J, et al. CANT1 mutation is also responsible for Desbuquois dysplasia, type 2 and Kim variant[J]. J Med Genet, 2011, 48(1): 32-37. DOI: 10.1136/jmg.2010.080226.
[8]FADEN M, ALZAHRANI F, ARAFAH D, et al. Mutation of CANT1 causes Desbuquois dysplasia[J]. Am J Med Genet A, 2010, 152A(5): 1157-1160. DOI: 10.1002/ajmg.a.33404.
[9]BALASUBRAMANIAN K, LI B, KRAKOW D, et al. MED resulting from recessively inherited mutations in the gene encoding calciumactivated nucleotidase CANT1[J]. Am J Med Genet A, 2017, 173(9): 2415-2421. DOI: 10.1002/ajmg.a.38349.
[10]SHAN L, ZHOU X, LIU X, et al. FOXK2 elicits massive transcription repression and suppresses the hypoxic response and breast cancer carcinogenesis[J]. Cancer Cell, 2016, 30(5): 708-722. DOI: 10.1016/j.ccell.2016.09.010.
[11]皮国成, 肖何, 罗佳, 等. Torin2对人非小细胞肺癌A549细胞生长和迁移的影响[J]. 第三军医大学学报, 2016, 38(19): 2138-2143. DOI:10.16016/j.10005404.201609065.
PI G C,XIAO H,LUO J,et al. Effects of Torin2 on cell proliferation and migration in human nonsmall lung cancer A549 cells[J]. J Third Mil Med Univ, 2016, 38(24): 2576-2579. DOI: 10.16016/j.10005404.201609065.
[12]TOMITA Y. Early renal cell cancer[J]. Int J Clin Oncol, 2006, 11(1): 22-27. DOI: 10.1007/s1014700505514.
[13]RAVAUD A, MOTZER R J, PANDHA H S, et al. Adjuvant sunitinib in highrisk renalcell carcinoma after nephrectomy[J]. N Engl J Med, 2016, 375(23): 2246-2254. DOI: 10.1056/NEJMoa1611406.
[14]GERHARDT J, STEINBRECH C, BUCHI O, et al. The androgenregulated calciumactivated nucleotidase 1 (CANT1) is commonly overexpressed in prostate cancer and is tumorbiologically relevant in vitro[J]. Am J Pathol, 2011, 178(4): 1847-1860. DOI: 10.1016/j.ajpath.2010.12.046.

更新日期/Last Update: 2018-03-22