跨膜蛋白40（TMEM40）的上调与恶性行为相关，并促进宫颈癌的肿瘤进展。

Upregulation of TMEM40 is associated with the malignant behavior and promotes tumor progression in cervical cancer.

作者信息

Zhang Zhen-Fei, Liu Fang, Zhang Han-Rong, Liu Bing, Zheng Shu-Qian, Ye Wan-Qian, Ding Jia-Nan, Zhou Ze-Jie, Luo Hui-Xian, Wu Fang, Guo Xuan-Min, Zhou Jue-Yu, Guo Yong-Hui

机构信息

Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, 253 Industrial Avenue Central, Guangzhou, 510280, People's Republic of China.

Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, People's Republic of China.

出版信息

Discov Oncol. 2023 Apr 13;14(1):43. doi: 10.1007/s12672-023-00648-9.

DOI:10.1007/s12672-023-00648-9

PMID:37052818

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10102277/

Abstract

OBJECTIVE

Recent studies indicated that transmembrane protein 40 (TMEM40) is associated with several types of cancers but is not clear in cervical cancer (CC). The study aimed to examine the role of TMEM40 in CC and related mechanisms.

METHODS

The expression of TMEM40 in CC tissues and cell lines was studied with western blot and real-time quantitative RT-PCR. The effect of TMEM40 on proliferation was evaluated by CCK-8, EdU and colony formation assay. The migration, invasion, cell cycle and apoptosis of CC cells were studied with wound healing, transwell assays and flow cytometry. Tumor growth was evaluated in vivo using a xenogenous subcutaneously implant model.

RESULTS

The results revealed that the TMEM40 elevation in CC tissues and cell lines was closely correlated with tumor size and lymph node metastasis in clinical patients. Upregulation of TMEM40 with OE-TMEM40 vector promoted the invasion, migration and proliferation, inhibited the apoptosis and led to distinct S cell cycle arrest in CC cell lines. Silencing TMEM40 with shRNA inhibited the invasion, migration and proliferation, promoted apoptosis and led to a G0/G1 cell cycle arrest in CC cell lines. Silence of TMEM40 downregulated the expression of c-MYC, Cyclin D1, matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-9 (MMP-9), but in contrast, activated p53 and several apoptosis related proteins such as p53, Caspase-3, Caspase-9 and PARP1. In addition, TMEM40 silencing dramatically decreased tumor growth in mice models.

CONCLUSION

The present study demonstrates that TMEM40 upregulation can be a potential prognostic biomarker and contribute to CC development.

摘要

目的

近期研究表明，跨膜蛋白40（TMEM40）与多种癌症相关，但在宫颈癌（CC）中的情况尚不清楚。本研究旨在探讨TMEM40在CC中的作用及相关机制。

方法

采用蛋白质免疫印迹法和实时定量逆转录聚合酶链反应研究TMEM40在CC组织和细胞系中的表达。通过细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）和集落形成试验评估TMEM40对增殖的影响。采用伤口愈合试验、Transwell试验和流式细胞术研究CC细胞的迁移、侵袭、细胞周期和凋亡。使用异种皮下植入模型在体内评估肿瘤生长。

结果

结果显示，CC组织和细胞系中TMEM40的升高与临床患者的肿瘤大小和淋巴结转移密切相关。用OE-TMEM40载体上调TMEM40可促进CC细胞系的侵袭、迁移和增殖，抑制凋亡并导致明显的S期细胞周期阻滞。用短发夹RNA（shRNA）沉默TMEM40可抑制CC细胞系的侵袭、迁移和增殖，促进凋亡并导致G0/G1期细胞周期阻滞。沉默TMEM40可下调c-MYC、细胞周期蛋白D1、基质金属蛋白酶-1（MMP-1）和基质金属蛋白酶-9（MMP-9）的表达，但相反，可激活p53和几种凋亡相关蛋白，如p53、半胱天冬酶-3（Caspase-3）、半胱天冬酶-9（Caspase-9）和聚（ADP-核糖）聚合酶1（PARP1）。此外，沉默TMEM40可显著降低小鼠模型中的肿瘤生长。