CircRNA_PLEKHM3 regulates epithelial mesenchymal transformation of cervical cancer cells through the miR-320/KLF4 axis
10.19405/j.cnki.issn1000-1492.2024.03.006
- Author:
Yanan Zhang
1
;
Ying Cui
1
;
Tianjiao Wang
1
;
Zhonglei Du
1
Author Information
1. Obstetrics and Gynecology Ward , Hongqi Hospital Afiliated to Mudanjiang Medical College , Mudanjiang 157000
- Publication Type:Journal Article
- Keywords:
circular RNA_PLEKHM3;
miRNA 320;
krüppel like factor 4;
cervical cancer cells;
epithelial mes enchymal transition;
migration;
invasion
- From:
Acta Universitatis Medicinalis Anhui
2024;59(3):403-412
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To investigate the role and mechanism of circular RNA containing pleck strin homology do main of Pleck strin homology domain family M member 3 ( circRNA_PLEKHM3) in regulating epithelial mesenchy mal transition (EMT) behavior in cervical cancer cells through the miR-320 and KLF4 .
Methods :The expression levels of circRNA_PLEKHM3 in cervical cancer cells Hela and CaSki were detected by real time quantitative PCR (qRT PCR) . RNA fluorescence in situ hybridization was used to determine the localization of circRNA_PLEKHM3 in human cervical cancer epithelial cells CaSki . Dual luciferase reporter gene experiments were conducted to inves tigate the targeting relationship between circRNA_PLEKHM3 and miR-320 , as well as the targeting relationship be tween miR-320 and KLF4 . CaSki cells were overexpressed with circRNA_PLEKHM3. Additionally, three groups were set up: overexpression of miR-320 on the basis of circRNA_PLEKHM3 overexpression , silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression , and silencing of KLF4 on the basis of miR-320 overexpression . qRT PCR was performed to detect the expression levels of miR-320 in CaSki . Western blot experiments were con ducted to determine the expression of KLF4 and EMT markers including E-cadherin , N-cadherin , Vimentin , MMP-2 , and MMP-9 in CaSki cells . Transwell assays were performed to measure cell migration and invasion .
Results:The expression of circRNA_PLEKHM3 decreased in Hela and CaSki cells (P < 0 05) , mainly localized in the cytoplasm . The dual luciferase reporter gene experiment demonstrated a targeting relationship between miR-320 and circRNA_PLEKHM3 , as well as between KLF4 and miR-320 . Overexpression of circRNA_PLEKHM3 inhibited the protein expression of miR-320 , Ncadherin , Vimentin , MMP-2 , and MMP-9 , up regulated the protein expression of E-cadherin, and reduced cell migration and invasion ( P < 0.05) . Overexpression of miR 320 or silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression both promoted the protein expression of miR-320, N-cad herin , Vimentin , MMP-2 , and MMP-9 , down regulated the protein expression of E-cadherin , and increased cell migration and invasion (P < 0.05) . However , silencing of KLF4 on the basis of miR-320 overexpression inhibited the protein expression of KLF4 , N-cadherin , Vimentin , MMP-2 , and MMP-9 , up regulated the protein expression of E-cadherin , and reduced cell migration and invasion ( P < 0.05) .
Conclusion :Overexpression of circRNA _PLEKHM3 regulates EMT in cervical cancer cells through the miR -320/KLF4 axis .
- Full text:202406261037007332环状RNA＿PLEKHM3...调控宫颈癌细胞上皮间质转化_张亚男.pdf
