Effect of HPV E6 on Proliferation，Invasion and Migration of Cervical Cancer Cells Through Rap1 Signaling Pathway

Xiangchuan QIN; Jinqiu LI; Xiaojing HUANG; Kuerban HUTUBIDING·; Hasim AXIANGU·

doi:10.12259/j.issn.2095-610X.S20240902

Volume 45 Issue 9

Sep. 2024

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Xiangchuan QIN, Jinqiu LI, Xiaojing HUANG, Kuerban HUTUBIDING·, Hasim AXIANGU·. Effect of HPV E6 on Proliferation，Invasion and Migration of Cervical Cancer Cells Through Rap1 Signaling Pathway[J]. Journal of Kunming Medical University, 2024, 45(9): 9-16. doi: 10.12259/j.issn.2095-610X.S20240902

Citation:

Xiangchuan QIN, Jinqiu LI, Xiaojing HUANG, Kuerban HUTUBIDING·, Hasim AXIANGU·. Effect of HPV E6 on Proliferation，Invasion and Migration of Cervical Cancer Cells Through Rap1 Signaling Pathway[J]. Journal of Kunming Medical University, 2024, 45(9): 9-16. doi: 10.12259/j.issn.2095-610X.S20240902

Citation:

Xiangchuan QIN, Jinqiu LI, Xiaojing HUANG, Kuerban HUTUBIDING·, Hasim AXIANGU·. Effect of HPV E6 on Proliferation，Invasion and Migration of Cervical Cancer Cells Through Rap1 Signaling Pathway[J]. Journal of Kunming Medical University, 2024, 45(9): 9-16. doi: 10.12259/j.issn.2095-610X.S20240902

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Effect of HPV E6 on Proliferation，Invasion and Migration of Cervical Cancer Cells Through Rap1 Signaling Pathway

doi: 10.12259/j.issn.2095-610X.S20240902

Basic Medicine College of Xinjiang Medical University/Xinjiang Key Laboratory of Molecular Biology of Endemic Diseases，Urumqi Xinjiang 836001，China

Received Date: 2024-05-12
Available Online: 2024-09-04
Publish Date: 2024-09-25

Abstract

Abstract

Objective To investigate the effect of human papillomavirus E6 protein （HPV E6） on the proliferation, invasion and migration of Hela cells of cervical cancer and its potential molecular mechanism. Methods Illumina Hiseq 4000 transcriptome sequencing technology was employed to identify the gene expression profiles of 12 cases featuring various degrees of cervical lesions and to screen for HPV-related differential genes. The biological function enrichment of the differential genes was analyzed using GO and KEGG Pathway. Western blotting was utilized to detect the expression levels of Rap and Rap1GAP proteins in normal cervical epithelial cells HCerEpic and cervical cancer cells Hela. The capabilities of proliferation, invasion, and migration were examined by plate cloning, Transwell, and scratch tests. Intracellular HPV E6 expression was down-regulated through sh-E6 lentivirus transfection and classified into the control group （Hela cells）, the no-load group （sh-NON）, and the sh-E6 group （with low expression of HPV E6）. The expression of HPV E6 was diminished, and the proliferation ability and cell viability of each group were determined by plate cloning and CCK-8 assay. Transwell and scratch assays were adopted to detect cell invasion and migration. Western blotting was conducted to detect the expression levels of E6, Rap1, Rap1GAP, CyclinD1, E-cadherin, and N-cadherin in cells of each group. Rap1 was overexpressed based on the sh-E6 group, and the expression levels of the Rap1 pathway and related proteins were measured by Western blot. Cell proliferation, invasion, and migration were evaluated respectively by plate cloning, Transwell assay, and scratch assay. Results The sequencing results indicated that, in contrast to normal cervical HPV-negative tissues, 340, 864 and 1036 differentially upregulated genes were respectively present in HPV-positive normal cervix, CIN and cervical cancer tissues. A total of 24 differential genes were identified across the 3 datasets. GO|KEGG enrichment analysis of the 24 differentially expressed genes primarily focused on the RAP1-related signaling pathway. Compared with HCerEpic cells, Rap1 expression was elevated and Rap1GAP expression was reduced in Hela cells （P < 0.01）. The proliferation, invasion and migration of the cells were enhanced （P < 0.01）. After down-regulating the expression of HPV E6, the proliferation, colony formation, invasion and migration of sh-E6 cells were decreased compared with the Hela group （P < 0.001）. Western blotting showed that the protein expressions of Rap1, CyclinD1 and N-cadherin in sh-E6 group were lower than those in Hela group, and the protein expressions of Rap1GAP and E-cadherin were higher than those in hela group （sh-E6/OE Rap1） were restored （P < 0.001）, and the cell proliferation, invasion and migration were restored （P < 0.01）. Conclusion During the development of cervical cancer, HPV E6 promotes the proliferation, invasion and migration of cervical cancer cells by activating Rap1 signaling pathway.
- HPV E6,
- Cervical cancer,
- Rap1 pathway,
- Proliferation,
- Invasion,
- Migration

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