Effect of miR-96-5p on Ishikawa Cells by Targeting Forkhead Box Transcription Factor O1
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2022.0407
- VernacularTitle:miR-96-5p靶向叉头状转录因子O1对Ishikawa细胞的作用
- Author:
Yan-qi FENG
1
;
E ZHANG
2
;
Lei ZHANG
1
;
Song-hua HAO
1
;
Ting-ting NI
3
Author Information
1. Department of Gynecology， Henan Medical College， Zhengzhou 451191， China
2. Department of Obstetrics and Gynecology， the First Affiliated Hospital of Henan University of Traditional Chinese Medicine， Zhengzhou 450000， China
3. Henan University of Traditional Chinese Medicine， Zhengzhou 450046， China
- Publication Type:Journal Article
- Keywords:
endometrial cancer;
miR-96-5p;
forkhead box transcription factor O1;
cyclin D1;
invasion
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(4):573-581
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the effect of microRNA-96-5p （miR-96-5p） on the proliferation and invasion of endometrial cancer cells by targeting forkhead box transcription factor O1 （FOXO1） genes. MethodsThe expressions of miR-96-5p and FOXO1 mRNA in normal and endometrial carcinoma tissues were detected by qRT-PCR， and FOXO1 protein expression was detected by Western blot. The relationship between miR-96-5p and FOXO1 expression and clinicopathological features of endometrial carcinoma was analyzed. pcDNA， pcDNA-FOXO1， inhibitor NC， miR-96-5p inhibitor， miR-96-5p mimic and miR-96-5p + FOXO1 were transfected into endometrial carcinoma Ishikawa cells， which were recorded as pcDNA group， FOXO1 group， inhibitor NC group， miR-96-5p inhibitor group， miR-96-5p group and miR-96-5p + FOXO1 group， respectively. Cells in stable transfection groups were taken， CCK-8 and Transwell chamber were employed to detect cell viability and invasion， and the expression levels of cyclin D1， cleaved PARP， p21 and Vimentin were measured by Western blot. TargetScan website was used to predict the targeting relationship between miR-96-5p and FOXO1， and dual luciferase reporter gene was used to detect cell luciferase activity. Ishikawa cells were divided into mimic NC group， miR-96-5p group， inhibitor NC group and miR-96-5p inhibitor group. The expressions of miR-96-5p and FOXO1 mRNA were detected by qRT-PCR， and the protein expression of FOXO1 was detected by Western blot. ResultsThe relative expression level of miR-96-5p in endometrial carcinoma tissues was up-regulated compared with that in normal tissues. On the contrary， the relative expression levels of FOXO1 mRNA and protein were down-regulated （P<0.01）. The high expression of miR-96-5p was positively correlated with pathological grade and clinical stage （P=0.034， P=0.010）， but not significantly correlated with age and menopause in endometrial carcinoma patients （P=0.370， P=0.166）. The low expression of FOXO1 was negatively correlated with pathological grade and clinical stage （P=0.023， P=0.007）， but not significantly correlated with the age and menopause in endometrial cancer patients （P=0.344， P=0.144）. After FOXO1 overexpression or inhibition of miR-96-5p expression， the viability of Ishikawa cells decreased significantly （P < 0.05）， the number of invasive cells decreased （P < 0.01）， the expression levels of cyclin D1 and Vimentin proteins decreased significantly， and the expression levels of cleaved PARP and p21 proteins increased significantly （P < 0.01）. Compared with the control group， the viability of Ishikawa cells was significantly increased （P < 0.05）， the number of invasive cells was increased （P < 0.01）， the expression levels of cyclin D1 and Vimentin were significantly increased， and the expression levels of cleaved PARP and p21 were significantly decreased in miR-96-5p group （P < 0.01）. Compared with the miR-96-5p group， overexpression of FOXO1 could significantly reverse the promotion of miR-96-5p on the viability and invasion of Ishikawa cells （P < 0.05）. miR-96-5p targeted and negatively regulated FOXO1 expression （P < 0.05）. ConclusionmiR-96-5p promotes the proliferation and invasion of endometrial cancer cells by targeting and negatively regulating FOXO1.
