Abstract
To investigate the effects of transmembrane protein 45A (TMEM45A) on biological characteristics and cisplatin (DDP) resistance of cervical cancer cells. TMEM45A in cervical cancer cells and normal cervical epithelial cells (HCerEpiC) were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. HPV genotypes were identified by multiplex PCR. SiHa and HeLa cells were divided into Blank, shCTL, shTMEM45A-1, and shTMEM45A-2 groups, followed by Cell Counting Kit-8 (CCK-8), EdU, Annexin V-FITC/PI staining, Wound healing, and Transwell invasion assays, as well as qRT-PCR and Western blotting. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) was employed to evaluate the impact of TMEM45A shRNA on cisplatin-resistant cervical cancer cells (SiHa/DDP and HeLa/DDP). Compared with HcerEpic cell, cervical cancer cells exhibited the upregulation of TMEM45A expression, especially in HPV-positive cell lines (CaSki, SiHa, HeLa). TMEM45A shRNA suppressed the proliferation of SiHa and HeLa cells, arrested cells at the S phase, and promoted cell apoptosis. TMEM45A shRNA inhibited the epithelial-mesenchymal transition (EMT), invasion, migration of SiHa and HeLa cells, accompanying by the downregulated Vimentin and N-cadherin with the upregulated E-cadherin. Moreover, SiHa/DDP and HeLa/DDP had higher TMEM45A expression than their parental SiHa and HeLa cells, respectively. And inhibiting TMEM45A can reduce the IC50 of SiHa/DDP cells and HeLa/DDP cells to cisplatin. Silencing TMEM45A can inhibit cell proliferation, invasion, migration and EMT, regulate cell cycle distribution, promote cell apoptosis, and reverse cisplatin resistance of HPV-positive cervical cancer cells, highlighting that inhibition of TMEM45A may be a therapeutic strategy for HPV-positive cervical cancer.
Similar content being viewed by others
References
Campo L, Zhang C, Breuer EK (2015) EMT-Inducing Molecular Factors in Gynecological Cancers BioMed Research International 2015:420891. https://doi.org/10.1155/2015/420891
Chen H, Tang X, Zhou B, Xu N, Zhou Z, Fang K, Wang Y (2017) BDE-47 and BDE-209 inhibit proliferation of Neuro-2a cells via inducing G1-phase arrest. Environ Toxicol Pharmacol 50:76–82. https://doi.org/10.1016/j.etap.2016.12.009
Flamant L et al (2012) TMEM45A is essential for hypoxia-induced chemoresistance in breast and liver cancer cells. BMC Cancer 12:391. https://doi.org/10.1186/1471-2407-12-391
Flanagan L et al (2016) Low levels of Caspase-3 predict favourable response to 5FU-based chemotherapy in advanced colorectal cancer: Caspase-3 inhibition as a therapeutic approach. Cell Death Dis 7:e2087. https://doi.org/10.1038/cddis.2016.7
Gaspar N et al (2009) Acquired resistance to 17-allylamino-17-demethoxygeldanamycin (17-AAG, tanespimycin) in glioblastoma cells. Can Res 69:1966–1975. https://doi.org/10.1158/0008-5472.CAN-08-3131
Gray P et al (2019) Occurrence of human papillomavirus (HPV) type replacement by sexual risk-taking behaviour group: Post-hoc analysis of a community randomized clinical trial up to 9 years after vaccination (IV). Int J Cancer 145:785–796. https://doi.org/10.1002/ijc.32189
Guo J, Chen L, Luo N, Yang W, Qu X, Cheng Z (2015) Inhibition of TMEM45A suppresses proliferation, induces cell cycle arrest and reduces cell invasion in human ovarian cancer cells. Oncol Rep 33:3124–3130. https://doi.org/10.3892/or.2015.3902
Hayez A et al (2014) High TMEM45A expression is correlated to epidermal keratinization. Exp Dermatol 23:339–344. https://doi.org/10.1111/exd.12403
Heselmeyer-Haddad K et al (2005) Genomic amplification of the human telomerase gene (TERC) in pap smears predicts the development of cervical cancer. Am J Pathol 166:1229–1238. https://doi.org/10.1016/S0002-9440(10)62341-3
Huo W et al (2020) Relevance research between the expression of p16(INK4a), Notch1, and hTERC genes: The development of HPV16-positive cervical cancer. J Clin Lab Anal 34:e23207. https://doi.org/10.1002/jcla.23207
Itoigawa Y et al (2015) TWEAK enhances TGF-beta-induced epithelial-mesenchymal transition in human bronchial epithelial cells. Respir Res 16:48. https://doi.org/10.1186/s12931-015-0207-5
Jing X, Yang F, Shao C, Wei K, Xie M, Shen H, Shu Y (2019) Role of hypoxia in cancer therapy by regulating the tumor microenvironment. Mol Cancer 18:157. https://doi.org/10.1186/s12943-019-1089-9
Lin BY, Wen JL, Zheng C, Lin LZ, Chen CZ, Qu JM (2020) Eva-1 homolog A promotes papillary thyroid cancer progression and epithelial-mesenchymal transition via the Hippo signalling pathway. J Cell Mol Med. https://doi.org/10.1111/jcmm.15909
Manawapat-Klopfer A et al (2016) TMEM45A, SERPINB5 and p16INK4A transcript levels are predictive for development of high-grade cervical lesions. Am J Cancer Res 6:1524–1536
Martin-Rendon E et al (2007) Transcriptional profiling of human cord blood CD133+ and cultured bone marrow mesenchymal stem cells in response to hypoxia. Stem Cells 25:1003–1012. https://doi.org/10.1634/stemcells.2006-0398
Modi A et al (2020) Benzothiazole derivative bearing amide moiety induces p53-mediated apoptosis in HPV16 positive cervical cancer cells. Invest New Drugs 38:934–945. https://doi.org/10.1007/s10637-019-00848-7
Park SB et al (2009) bFGF enhances the IGFs-mediated pluripotent and differentiation potentials in multipotent stem cells. Growth Factors 27:425–437. https://doi.org/10.3109/08977190903289875
Sample KM (2020) DNA repair gene expression is associated with differential prognosis between HPV16 and HPV18 positive cervical cancer patients following radiation therapy. Sci Rep 10:2774. https://doi.org/10.1038/s41598-020-59383-8
Schmit K et al (2019) Characterization of the role of TMEM45A in cancer cell sensitivity to cisplatin. Cell Death Dis 10:919. https://doi.org/10.1038/s41419-019-2088-x
Schmit K, Michiels C (2018) TMEM Proteins in Cancer: A Review Front Pharmacol 9:1345. https://doi.org/10.3389/fphar.2018.01345
Sun W et al (2015) Knockdown of TMEM45A inhibits the proliferation, migration and invasion of glioma cells. Int J Clin Exp Pathol 8:12657–12667
Wrzesinski T et al (2015) Expression of pre-selected TMEMs with predicted ER localization as potential classifiers of ccRCC tumors. BMC Cancer 15:518. https://doi.org/10.1186/s12885-015-1530-4
Xia W, Li Y, Wu Z, Wang Y, Xing N, Yang W, Wu S (2020) Transcription factor YY1 mediates epithelial-mesenchymal transition through the TGFbeta signaling pathway in bladder cancer. Med Oncol 37:93. https://doi.org/10.1007/s12032-020-01414-5
Yang YC et al (2001) Frequent gain of copy number on the long arm of chromosome 3 in human cervical adenocarcinoma. Cancer Genet Cytogenet 131:48–53. https://doi.org/10.1016/s0165-4608(01)00510-6
Zhang L, Wu F, Zhao J (2020) Transmembrane protein 45A regulates the proliferation, migration, and invasion of glioma cells through nuclear factor kappa-B. Anticancer Drugs 31:900–907. https://doi.org/10.1097/CAD.0000000000000890
Zhu M, Jiang B, Yan D, Wang X, Ge H, Sun Y (2020) Knockdown of TMEM45A overcomes multidrug resistance and epithelial-mesenchymal transition in human colorectal cancer cells through inhibition of TGF-beta signalling pathway. Clin Exp Pharmacol Physiol 47:503–516. https://doi.org/10.1111/1440-1681.13220
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Authors declared no conflicts of interest in this article.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Liu, Y., Liu, L. & Mou, ZX. TMEM45A Affects Proliferation, Apoptosis, Epithelial-Mesenchymal Transition, Migration, Invasion and Cisplatin Resistance of HPV-Positive Cervical Cancer Cell Lines. Biochem Genet 60, 173–190 (2022). https://doi.org/10.1007/s10528-021-10094-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10528-021-10094-3