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CDKN2A(p16INK4A/p14ARF)过表达对人黑色素瘤A375细胞增殖和迁移的影响

Effects of CDKN2A (p16INK4A/p14ARF) Over-Expression on Proliferation and Migration of Human Melanoma A375 Cells.

作者信息

Bai Ming, Yu Nan-Ze, Long Fei, Feng Cheng, Wang Xiao-Jun

机构信息

Division of Plastic and Reconstructive Surgery, Peking Union Medical College Hospital, Beijing, P.R. China.

出版信息

Cell Physiol Biochem. 2016;40(6):1367-1376. doi: 10.1159/000453189. Epub 2016 Dec 19.

Abstract

OBJECTIVE

This study aims to investigate the effects of CDKN2A (p16INK4A/p14ARF) over-expression on the proliferation and migration of human melanoma A375 cells.

METHODS

Melanoma tissues and pigmented nevi tissues were collected. Human melanoma A375 cells were transfected by CDKN2A (p16INK4A) and CDKN2A (p14ARF) over-expressing vectors and then assigned into blank, negative control (NC), p16INK4A and p14ARF groups. The expression of CDKN2A (p16INK4A) and CDKN2A (p14ARF) mRNA and protein was detected by qRT-PCR and Western blotting. CCK-8, flow cytometry and Transwell assays were applied to observe cell proliferation, the cell cycle and apoptosis, and migration and invasion, respectively. The model of subcutaneous xenografts in nude mice was established to measure cell growth in vivo.

RESULTS

Compared with pigmented nevi tissues, CDKN2A (p16INK4A) and CDKN2A (p14ARF) mRNA and protein expression were significantly decreased in melanoma tissues. CDKN2A (p16INK4A) and CDKN2A (p14ARF) over-expression inhibited proliferation, migration, invasion and progression from G0/G1 to S phase of A375 cells and xenograft tumor growth, but promoted apoptosis.

CONCLUSION

Our study demonstrated that over-expression of CDKN2A (p16INK4A) and CDKN2A (p14ARF) suppressed proliferation and migration of human melanoma A375 cells.

摘要

目的

本研究旨在探讨CDKN2A(p16INK4A/p14ARF)过表达对人黑色素瘤A375细胞增殖和迁移的影响。

方法

收集黑色素瘤组织和色素痣组织。将人黑色素瘤A375细胞用CDKN2A(p16INK4A)和CDKN2A(p14ARF)过表达载体转染,然后分为空白组、阴性对照组(NC)、p16INK4A组和p14ARF组。通过qRT-PCR和蛋白质免疫印迹法检测CDKN2A(p16INK4A)和CDKN2A(p14ARF)mRNA和蛋白的表达。分别采用CCK-8法、流式细胞术和Transwell实验观察细胞增殖、细胞周期和凋亡以及迁移和侵袭情况。建立裸鼠皮下异种移植模型以测量体内细胞生长情况。

结果

与色素痣组织相比,黑色素瘤组织中CDKN2A(p16INK4A)和CDKN2A(p14ARF)mRNA和蛋白表达明显降低。CDKN2A(p16INK4A)和CDKN2A(p14ARF)过表达抑制了A375细胞的增殖、迁移、侵袭以及从G0/G1期到S期的进程和异种移植瘤生长,但促进了细胞凋亡。

结论

我们的研究表明,CDKN2A(p16INK4A)和CDKN2A(p14ARF)过表达抑制了人黑色素瘤A375细胞的增殖和迁移。

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