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. 2024 Nov;38(11):1017-1023.
doi: 10.13201/j.issn.2096-7993.2024.11.005.

[Effect of phosphorylated HSP27 on the proliferation and metastasis of nasopharyngeal carcinoma and its mechanism]

[Article in Chinese]
Shan Bi  1   2 , Huajie Wu  1 , Han Fan  1 , Longmei Wan  1 , Guoping Li  1
Affiliations

[Effect of phosphorylated HSP27 on the proliferation and metastasis of nasopharyngeal carcinoma and its mechanism]

[Article in Chinese]
Shan Bi et al. Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2024 Nov.

Abstract
in English, Chinese

Objective:To investigate the effect of phosphorylated HSP27 on the proliferation and metastasis of nasopharyngeal carcinoma and its molecular mechanism. Methods:①Western blot assay was used to detect the expression levels of HSP27 and p-HSP27 in CNE1 and CNE2 cells of nasopharyngeal carcinoma. Inhibited the phosphorylation of HSP27, Transwell assay detected the metastasis ability of nasopharyngeal carcinoma cells. ②Nasopharyngeal carcinoma cell lines（CNE2-OE1 and CNE2-OE2） overexpressing phosphorylated and dephosphorylated mutants of HSP27 were synthesized, empty vector transfected CNE2 cells（CNE2-NC） were used as controls. The proliferation ability of the three groups of cells was detected by CCK8, and the expression levels of CyclinD1, Bax and Bcl-2 were detected by Western blot. Transwell was used to detect the migration and invasion ability of cells, and Western blot was used to detect the expression levels of E-cadherin, Vimentin, MMP2 and MMP9. Results:The expression level of HSP27 in CNE2 was higher than that of CNE1 cells, while the expression level of p-HSP27 was opposite. After inhibition of HSP27 phosphorylation, the invasion and migration ability of CNE1 cells decreased significantly, with no significant change in CNE2 cells. Compared with CNE2-NC, the growth rate of CNE2-OE1 decreased, and the growth rate of CNE2-OE2 increased. The expression level of CyclinD1 was down-regulated in CNE2-OE1 and higher in CNE2-OE2. The expression level of Bax in CNE2-OE1 was increased, while the expression of Bcl-2 was decreased. There was no significant change in the expression of Bax and Bcl-2 in CNE2-OE2. Compared with CNE2-NC, the migration ability of CNE2-OE1 was enhanced and the invasion ability was weakened, while the migration ability of CNE2-OE2 was weakened and the invasion ability was enhanced. There was no significant difference in the expression levels of E-cadherin was decreased in CNE2-OE1 and increased in CNE2-OE2. There was no significant difference in the expression levels of Vimentin among the three groups. The expression levels of MMP2 and MMP9 were up-regulated in CNE2-OE2, and slightly down-regulated in CNE2-OE1. Conclusion:HSP27 and p-HSP27 were differentially expressed in different nasopharyngeal carcinoma cells, and the metastasis ability of nasopharyngeal carcinoma was not only related to the expression level of HSP27, but also related to the level of p-HSP27. The p-HSP27 inhibited CNE 2 cell proliferation and promoted their apoptosis. As p-HSP27 plays different roles in different stages of nasopharyngeal carcinoma metastasis, it can increase the migration ability of CNE2 cells and reduce its invasion ability. p-HSP27 may induce EMT changes in CNE2 by down-regulating E-cadherin, thus promoting CNE2 migration, and may inhibit CNE2 invasion by down-regulating MMPs expression.

目的：探讨磷酸化HSP27（p-HSP27）对鼻咽癌增殖和转移的影响及其分子机制。 方法：①Western blot检测鼻咽癌CNE1、CNE2细胞中HSP27和p-HSP27的表达水平；抑制HSP27磷酸化，Transwell检测细胞的迁移、侵袭能力。②合成过表达HSP27磷酸化和去磷酸化突变体的鼻咽癌细胞株（CNE2-OE1和CNE2-OE2），以转染空载体的CNE2细胞（CNE2-NC）为对照：CCK8检测3组细胞的增殖能力，Western blot检测CyclinD1、Bax和Bcl-2的表达水平；Transwell检测细胞的迁移、侵袭能力，Western blot检测E-cadherin、Vimentin、MMP2、MMP9的表达水平。 结果：①HSP27在CNE2中的表达高于CNE1，p-HSP27的表达则相反；抑制HSP27磷酸化以后，CNE1的侵袭、迁移能力显著降低，CNE2无明显变化。②与CNE2-NC比较：CNE2-OE1生长速度降低，CNE2-OE2生长速度加快；CyclinD1在CNE2-OE1中表达下调，在CNE2-OE2中表达较高；CNE2-OE1中Bax的表达升高，Bcl-2的表达降低，两者在CNE2-OE2中的表达无明显变化。③与CNE2-NC比较：CNE2-OE1的迁移能力增强、侵袭能力减弱，CNE2-OE2的迁移能力减弱而侵袭能力增强；E-cadherin在CNE2-OE1中表达减少、在CNE2-OE2中表达增加，Vimentin在3组细胞中的表达差异无统计学意义；MMP2和MMP9在CNE2-OE2中表达上调，在CNE2-OE1中表达略下调。 结论：①HSP27和p-HSP27在不同鼻咽癌细胞中呈差异性表达，鼻咽癌的转移能力不仅与HSP27、也与p-HSP27的表达水平有关。②p-HSP27可抑制CNE2细胞增殖，促进其凋亡。③p-HSP27在鼻咽癌转移的不同环节发挥不同作用，其可增加CNE2细胞的迁移能力、降低其侵袭能力；p-HSP27可能通过下调E-cadherin使CNE2细胞发生EMT改变进而促进CNE2细胞迁移，通过下调MMPs的表达抑制CNE2细胞侵袭。.

Keywords: HSP27; metastasis; nasopharyngeal carcinoma; phosphorylation; proliferation.

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Conflict of interest statement

The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose.

Figures

图 1
图 1
HSP27和p-HSP27在CNE1、CNE2细胞中的表达水平
图 2
图 2
抑制HSP27磷酸化后，CNE1和CNE2细胞迁移、侵袭能力的改变
图 3
图 3
慢病毒转染荧光图
图 4
图 4
CNE2-NC、CNE2-OE1、CNE2-OE2细胞的增殖能力
图 5
图 5
CyclinD1在CNE2-NC、CNE2-OE1、CNE2-OE2细胞中的表达水平
图 6
图 6
HSP27磷酸化对Bax和Bcl-2表达量的影响
图 7
图 7
HSP27磷酸化对鼻咽癌细胞迁移能力的影响
图 8
图 8
HSP27磷酸化对鼻咽癌细胞侵袭能力的影响
图 9
图 9
HSP27磷酸化对EMT相关蛋白表达的影响
图 10
图 10
HSP27磷酸化对MMPs表达量的影响
See this image and copyright information in PMC

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