IMR Press / EJGO / Volume 41 / Issue 5 / DOI: 10.31083/j.ejgo.2020.05.5129
Open Access Original Research
Effect of CD59-targeted multi-directional intervention on transmembrane apoptotic signal transduction of cervical cancer HeLa cells
Tao Wei1,2,†Mei-Hua Gao1,†,*Ying Li3
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1 Department of Immunology, Medical College of Qingdao University, Qingdao 266071, P. R. China
2 Department of Medical Affairs, Hospital office, Qingdao Women and Children's Hospital, Qingdao University, Qingdao, 266000, P. R. China
3 Department of Blood Transfusion, The affiliated hospital of Qingdao university, Qingdao 266000, P. R. China

†Contributed equally.

Eur. J. Gynaecol. Oncol. 2020 , 41(5), 713–719; https://doi.org/10.31083/j.ejgo.2020.05.5129
Submitted: 2 July 2019 | Accepted: 18 September 2019 | Published: 15 October 2020
Abstract

Objective: With GPI-anchored protein CD59 as the target, the effect of the abnormal expression of the CD59 gene on the proliferation and apoptosis of cervical cancer HeLa cells was studied to investigate the mechanism of the malignant proliferation of cervical cancer cells. Materials and Methods: Recombinant plasmid pSUPER-siCD59 (CD59 RNA interference) was transfected into HeLa cells via liposome transfection. The plasmid expressed green fluorescent proteins that enabled the observation of the transfection efficiency and was separately transfected. The co-transfection of a pIRES-WTCD59 (wild-type CD59) plasmid, pIRES-MCD59 (with CD59W40 active site mutation) plasmid, and pLeGFP plasmid helped the direct observation of the transfection efficiency, which was examined via an inverted fluorescence microscope at 24, 48, and 72 hours after transfection. Then, the stable transfected cell lines were screened by G418. Western blot, RT-PCR, and immunofluorescence were used to detect the gene and protein expression of CD59 in the stably transfected HeLa cell lines, and the differences were evaluated. The short peptide seal specific to CD59 was exerted on the HeLa cells. Cell proliferation was detected by MTT colorimetric assay, and apoptosis was detected by TUNEL and flow cytometry. Results: The proliferation activity of the HeLa cells with CD59RNAi, CD59W40 active site mutations, and the short peptide seal specific to CD59 significantly decreased. In particular, the proliferation activity of the HeLa cells treated with a high dose of the short peptide seal specific to CD59 was significantly reduced, while the proliferation activity of the HeLa cells with a high expression of CD59 was significantly enhanced. Conclusion: As an inhibitory regulatory protein in the terminal stage of complement regulation, CD59 can promote the proliferation of cervical cancer HeLa cells via inhibiting the dissolution effect of the complement on the cells, promoting transmembrane activation signal transduction, and inhibiting apoptotic signal transduction. Thus, this study provides a new avenue for the clinical treatment of cervical cancer.

Keywords
CD59 knockout
Short-peptide seal
RNAi
Hela cell
Proliferation
Apoptosis
Figures
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