MicroRNA 95 Can Promote HCC Proliferation And Migration By Activating Akt/GSK3?/?-catenin Signaling Through ST7L

Objective: Hepatocellular carcinoma(HCC)is a kind of malignant tumor which threatens human life.The study on the molecular mechanism of primary liver cancer is a hot topic in China and all over the world.Many micro RNAs(mi RNA)have been proved to play an important role in the development of HCC.We selected micro RNA 95,and many studies have shown that mi R-95 can promote the growth of many tumors,such as colon cancer,liver cancer,breast cancer,prostate cancer,pancreatic cancer and non-small cell lung cancer.On the basis of previous studies,our study focuses on the role of Micro RNA 95 in the occurrence and development of HCC.Methods: Mi RNA high-throughput sequencing and qRT-PCR were used to detect the expression of mi R-95 in hepatocellular carcinoma tissues and normal liver tissues;CCK8 assay and clone formation assay were used to study the function of mi R-95 and ST7 L on the proliferation of HCC in vitro;Wound-healing assays and Transwell invasion assays were used to study the function of mi R-95 and ST7 L on the migration of HCC in vitro;HCC xenograft mouse model were used to study the effect of mi R-95 and ST7 L on HCC proliferation and migration in vivo;q RT-PCR assays were used tol study m RNA level of downstream genes of mi R-95/ST7L/Akt/ GSK3?/?-catenin signaling;Western-blot assays were used to study the related protein in mi R-95/ST7L/Akt/GSK3?/?-catenin signaling and the protein level of mi R-95/ST7L/Akt/GSK3?/?-catenin signaling downstream genes;immunofluorescence experiments were performed to study the co localization in cells of ST7 L and AKT protein;immunohistochemical assays were used to detect o related protein levels in HCC tissues and nude mice HCC tissues;CO-IP assays were used to study the interaction between ST7 L and AKT protein;SPSS were used to statistically analyze the relationship between mi R-95 HCC and the clinicopathological factors.Results: Results of histological assays showed that the expression of mi R-95 in hepatocellular carcinoma tissues was significantly higher than that in normal liver tissue,the expression level of mi R-95 was closely related to tumor size and tumor number;CCK8 and clone formation experiment showed that mi R-95 could promote the proliferation of HCC in vitro;Wound-healing assays and Transwell invasion assays showed that mi R-95 can promote the migration of HCC in in vitro;xenograft mouse model assays showed that mi R-95 can promote HCC proliferation and migration in vivo;dual luciferase reporter gene assay showed that ST7 L is a direct target of mi R-95;immunohistochemical results suggest that the expression level of ST7 L protein in HCC tissues was negatively correlated with the the expression level of mi R-95;rescue experimental results show that ST7 L can reverse the proliferation and migration of HCC induced by mi R-95 in vitro and in vivo.Mechanism study showed that mi R-95 can regulated Akt/GSK3?/?-catenin signaling pathway by its target ST7 L,inducing the related protein level changes of Akt/ GSK3?/?-catenin signaling and the m RNA and protein changes of downstream genes;immunofluorescence experiments showed that ST7 L and AKT proteins have the same cellular localization.CO-IPassays showed that ST7 L can interact with carboxyl terminal of AKT and induce Akt(Ser473)phosphorylation.Conclusion: the study shows that in human HCC tissues,the expression of mi R-95 in hepatocellular carcinoma tissues was significantly higher than that in normal liver tissue,the expression level of mi R-95 was closely related to tumor size and tumor number;Functional studies show that mi R-95 can promote the proliferation and migration of HCC in vitro and in vivo;mechanism research shows that ST7 L can interact with carboxy terminal of AKT where Akt(Ser473)lacated to inhibit phosphorylation of Akt(Ser473).mi R-95 can promote the proliferation and migration in vitro and in vivo by regulating Akt/GSK3?/?-catenin signaling axis in HCC through its direct target ST7L.