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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
Silencing calcium-activated nucleotidase 1 inhibits proliferation and migration of clear cell renal cell carcinoma 769-P cells
- 第三军医大学学报 2018年40卷第6期 页码：473-478
- 作者机构：
400037 重庆，陆军军医大学(第三军医大学)第二附属医院泌尿外科
- 作者简介：
[ 李龙坤，E-mail: lilongk@hotmail.com LI Longkun, E-mail: lilongk@hotmail.com
- 基金信息：国家自然科学基金面上项目(81470989)
DOI：10.16016/j.1000-5404.201711057
中图分类号： R394.3;R730.23;R737.11收稿：2017-11-08，
修回：2017-12-13，
纸质出版：2018-03-30
- 稿件说明：
移动端阅览
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(6):473-478.
Xin LIU, Xiaobing LIU, Qian LIU, et al. Silencing calcium-activated nucleotidase 1 inhibits proliferation and migration of clear cell renal cell carcinoma 769-P cells[J]. Journal of Third Military Medical University, 2018, 40(6): 473-478.
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(6):473-478. DOI： 10.16016/j.1000-5404.201711057.
Xin LIU, Xiaobing LIU, Qian LIU, et al. Silencing calcium-activated nucleotidase 1 inhibits proliferation and migration of clear cell renal cell carcinoma 769-P cells[J]. Journal of Third Military Medical University, 2018, 40(6): 473-478. DOI： 10.16016/j.1000-5404.201711057.
摘要
目的
2
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
2
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
2
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
2
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
2
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
2
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
2
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
2
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.
关键词
Keywords
references
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