PTEN C末端对鼻咽癌细胞迁移及增殖能力的影响

doi:10.3969/j.issn.1004-616x.2022.06.005

摘要/Abstract

摘要： 目的：研究PTEN C末端对鼻咽癌细胞迁移及增殖能力的影响。方法：采用荧光定量PCR实验检测PTEN mRNA在鼻咽癌细胞株HONE1、SUNE1、CNE1、CNE2、5-8F和鼻咽上皮细胞NP69中的表达情况。选择5-8F和HONE1细胞，用含有TGF-β1(5ng/mL)的RPMI 1640培养基进行细胞培养，分别瞬时转染阴性对照(NC)、PTEN WT(野生型)和缺乏C末端结构的PTEN 1-353(突变体)质粒，并采用Western blot实验验证转染效率。转染成功后48h，分别采用Transwell实验、CCK-8法细胞增殖实验检测PTEN WT和PTEN 1-353对鼻咽癌细胞株5-8F和HONE1细胞迁移、增殖的影响。结果：与NP69细胞相比，PTEN mRNA在鼻咽癌细胞中的表达水平明显降低(P<0.01)。与转染NC组比较，PTEN WT和PTEN 1-353均可以抑制TGF-β1诱导的鼻咽癌细胞的迁移(P<0.01)，但此二组之间差异无统计学意义(P>0.05)；PTEN WT和PTEN 1-353均可抑制TGF-β1诱导的鼻咽癌细胞增殖(P<0.05)，且相比于PTEN WT，PTEN 1-353对鼻咽癌细胞增殖的抑制能力降低(P<0.01)。结论：PTEN C末端未参与鼻咽癌细胞的迁移过程，但可能与鼻咽癌细胞的增殖有关。

关键词: PTEN C末端, 细胞迁移, 细胞增殖, 鼻咽癌

Abstract: OBJECTIVE:To elucidate functions of the PTEN C-tail in nasopharyngeal carcinoma (NPC). METHODS:Fluorescence quantitative PCR was used to detect expressions of PTEN in NPC cell lines and nasopharyngeal epithelial cell lines,NP69.5-8F and HONE1. Cells were cultured with 5 ng/mL TGF-β1. Negative control (NC),PTEN WT(wild-type) and PTEN 1-353 (lacking C-tail structure of PTEN) plasmids were over-expressed in the cells. Western blot were used to verify their transient efficiency. After 48 h,effects of PTEN WT and PTEN 1-353 on metastasis abilities were verified by Transwell experiments. Effect of PTEN WT and PTEN 1-353 on proliferation capacities were verified by CCK-8 assays. RESULTS:Compared with NP69,expressions of PTEN in NPC cells were reduced. Compared with the NC group,the migration abilities of TGF-β1-induced NPC cells were inhibited by PTEN WT and PTEN 1-353 (P<0.01),but there was no statistical difference between PTEN WT and PTEN 1-353 (P>0.05). Compared with the NC group,the proliferation abilities of TGF-β1-induced NPC cells were inhibited by PTEN WT and PTEN 1-353 (P<0.05). Interestingly,the inhibition of PTEN 1-353 was weaker-than PTEN WT(P<0.05). CONCLUSION:The PTEN C-tail was not involved in the regulation of metastasis of NPC cells. However,the PTEN C-tail could be involved in regulating the proliferation ability of NPC cells.

Key words: PTEN C-tail, cell migration, cell proliferation, nasopharyngeal carcinoma

中图分类号:

张军军, 梁乐平. PTEN C末端对鼻咽癌细胞迁移及增殖能力的影响[J]. 癌变·畸变·突变, 2022, 34(6): 434-438.

ZHANG Junjun, LIANG Leping. Effect of PTEN C-tail on migration and proliferation of nasopharyngeal carcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(6): 434-438.

参考文献

[1] CHEN C Y, CHEN J Y, HE L N, et al. PTEN:tumor suppressor and metabolic regulator[J]. Front Endocrinol (Lausanne), 2018, 9:338.
[2] STUMPF M, DEN HERTOG J. Differential requirement for pten lipid and protein phosphatase activity during zebrafish embryonic development[J]. PLoS One, 2016, 11(2):e0148508.
[3] VAZQUEZ F, RAMASWAMY S, NAKAMURA N, et al. Phosphorylation of the PTEN tail regulates protein stability and function[J]. Mol Cell Biol, 2000, 20(14):5010-5018.
[4] DEMPSEY D R, VIENNET T, IWASE R, et al. The structural basis of PTEN regulation by multi-site phosphorylation[J]. Nat Struct Mol Biol, 2021, 28(10):858-868.
[5] CORDIER F, CHAFFOTTE A, TERRIEN E, et al. Ordered phosphorylation events in two independent cascades of the PTEN C-tail revealed by NMR[J]. J Am Chem Soc, 2012, 134(50):20533-20543.
[6] WANG L Z, SANG J Z, ZHANG Y M, et al. Circular RNA ITCH attenuates the progression of nasopharyngeal carcinoma by inducing PTEN upregulation via miR-214[J]. J Gene Med, 2022, 24(1):e3391.
[7] 薛慧, 孟爱凤, 徐德静, 等. 鼻咽癌患者放疗期间医院感染发病率及影响因素[J]. 中国感染控制杂志, 2016, 15(7):481-483.
[8] 韩继波, 黄茂凌, 申丽君, 等. miR-214通过PTEN调控AKT信号通路抑制鼻咽癌细胞凋亡[J]. 肿瘤防治研究, 2020, 47(10):734-739.
[9] LIN C B, QIAN P F, ZHANG Y, et al. Plac1 promotes nasopharyngeal carcinoma cells proliferation, migration and invasion via Furin/NICD/PTEN pathway[J]. Tissue Cell, 2021, 69:101480.
[10] LIN J J, FAN J J, GE X J, et al. PAG1 stimulates proliferation and metastasis of nasopharyngeal carcinoma through downregulating PTEN[J]. Eur Rev Med Pharmacol Sci, 2020, 24(21):11096-11104.
[11] YAO L Z, ZHU Y L, LIU J J. Inhibition of PTEN gene expression by small interfering RNA on PI3K/Akt/FoxO3a signaling pathway in human nasopharyngeal carcinoma[J]. Technol Cancer Res Treat, 2020, 19:1533033820917959.
[12] 何雯, 杨森, 王琦, 等. circPVT1对鼻咽癌细胞增殖、转移和凋亡的影响及作用机制研究[J]. 国际检验医学杂志, 2022, 43(6):672-677, 682.
[13] 杨时平, 马骏, 应小芳, 等. 抑癌基因PTEN在鼻咽癌中的突变及表达[J]. 解放军医学杂志, 2009, 34(7):868-870, 880.
[14] 程金妹, 谢振宇, 林勤, 等. 抑癌基因PTEN在鼻咽癌中的表达及突变[J]. 山东大学耳鼻喉眼学报, 2008, 22(5):405-407.
[15] CORDIER F, CHAFFOTTE A, TERRIEN E, et al. Ordered phosphorylation events in two independent cascades of the PTEN C-tail revealed by NMR[J]. J Am Chem Soc, 2012, 134(50):20533-20543.
[16] VAZQUEZ F, RAMASWAMY S, NAKAMURA N, et al. Phosphorylation of the PTEN tail regulates protein stability and function[J]. Mol Cell Biol, 2000, 20(14):5010-5018.
[17] DEMPSEY D R, VIENNET T, IWASE R, et al. The structural basis of PTEN regulation by multi-site phosphorylation[J]. Nat Struct Mol Biol, 2021, 28(10):858-868.