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Research
Qin Wang
Sun Yat-Sen University
Corresponding Author
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This work is licensed under a CC BY 4.0 License. Read Full License
Background
The roles of gap junction and its components, connexins in autophagy of cervical cancer cells are rarely investigated. Our previous study demonstrated that connexin32 (Cx32) exerted an anti-apoptotic effect on cervical cancer (CaCx). However, as an important regulator of apoptosis, whether the autophagy is involved in the function of Cx32 on cervical cancer cells are not well defined.
Methods
The expression of Connexin32 and the autophagy-associated protein LC3-Ⅱ in paracancerous cervical tissues (n = 30) and cervical cancer (n = 50) tissues were determined by Western blotting. Forty-five cervical cancer specimens were used to evaluate the clinical relevance of Connexin32 and LC3-Ⅱ. The effect of Connexin32 on autophagy was examined by detecting the change of LC3-Ⅱ using Western blotting, transfection with enhanced green fluorescent protein-LC3 plasmid and transmission electron microscopy analysis. The mechanism of Connexin32-mediated autophagy was assessed by Western blotting and flow cytometry.
Results
Both Connexin32 and LC3-Ⅱ are upregulated in cervical cancer tissues compared to those in paracancerous cervical tissues. Overexpression of Connexin32 significantly enhanced autophagy in HeLa-Connexin32 cells, whereas knockdown of Connexin32 suppressed autophagy in C-33A cells. Connexin32 inhibited apoptosis of cervical cancer cells by promoting autophagy. Moreover, Connexin32 triggered autophagy by activation of the AMP-activated protein kinase signalling, which was gap junction-independent.
Conclusion
Connexin32 exerts its anti-apoptotic effect by activating autophagy through the AMPK pathway in cervical cancer. This finding suggests that Connexin32 is a potential biomarker and a new therapeutic target for cervical cancer.
Trial registration
The study was approved by the Research Committee of Ethics of the Affiliated Cancer Hospital of Xinjiang Medical University (K-201354).
Keywords
Connexin32, Cervical carcinoma, Autophagy, Apoptosis, AMP-activated protein kinase
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This is a preprint. It has not completed peer review.
Research
Qin Wang
Sun Yat-Sen University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 22 Mar, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
The roles of gap junction and its components, connexins in autophagy of cervical cancer cells are rarely investigated. Our previous study demonstrated that connexin32 (Cx32) exerted an anti-apoptotic effect on cervical cancer (CaCx). However, as an important regulator of apoptosis, whether the autophagy is involved in the function of Cx32 on cervical cancer cells are not well defined.
Methods
The expression of Connexin32 and the autophagy-associated protein LC3-Ⅱ in paracancerous cervical tissues (n = 30) and cervical cancer (n = 50) tissues were determined by Western blotting. Forty-five cervical cancer specimens were used to evaluate the clinical relevance of Connexin32 and LC3-Ⅱ. The effect of Connexin32 on autophagy was examined by detecting the change of LC3-Ⅱ using Western blotting, transfection with enhanced green fluorescent protein-LC3 plasmid and transmission electron microscopy analysis. The mechanism of Connexin32-mediated autophagy was assessed by Western blotting and flow cytometry.
Results
Both Connexin32 and LC3-Ⅱ are upregulated in cervical cancer tissues compared to those in paracancerous cervical tissues. Overexpression of Connexin32 significantly enhanced autophagy in HeLa-Connexin32 cells, whereas knockdown of Connexin32 suppressed autophagy in C-33A cells. Connexin32 inhibited apoptosis of cervical cancer cells by promoting autophagy. Moreover, Connexin32 triggered autophagy by activation of the AMP-activated protein kinase signalling, which was gap junction-independent.
Conclusion
Connexin32 exerts its anti-apoptotic effect by activating autophagy through the AMPK pathway in cervical cancer. This finding suggests that Connexin32 is a potential biomarker and a new therapeutic target for cervical cancer.
Trial registration
The study was approved by the Research Committee of Ethics of the Affiliated Cancer Hospital of Xinjiang Medical University (K-201354).
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
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