MiR-27b Regulates The Role Of SMAD1 In Osteosarcoma Cell Proliferation,Migration,Invasion And Apoptosis

Objective:Osteosarcoma is a malignant tumor that often occurs in children and adolescents.Clinical symptoms often manifest as bone,local masses,and joint pain.Although early detection and timely treatment have greatly improved the survival rate of patients,the mortality rate is still relatively high.High and prone to lung metastases.Neoadjuvant chemoradiotherapy combined with limb salvage is currently the main method for the treatment of osteosarcoma.However,radiotherapy and chemotherapy have a large toxic side effect on patients.Postoperative bone tissue reconstruction is required.Patients with metastases have high recurrence after surgery.The current treatment methods for osteosarcoma are neoadjuvant chemotherapy and radiation therapy combined with limb salvage surgery.Since chemotherapy drugs and radiation therapy seriously harm patients'bodies,adjuvant treatments such as postoperative bone tissue reconstruction can slow down the effects of drug toxic and side effects.This article seeks and validates miR-27b-3p and the target gene SMAD1,studies the effects of MiR-27b-3p on the proliferation,invasion,and migration of osteosarcoma cells,and provides a theoretical basis for targeted therapy of osteosarcoma.Methods:(1)After querying through PITA Sites and miRnada Sites,analyze and compare the sequence of miR-27b-3p gene and SMAD1.QRT-PCR experiments were used to measure the expression of miR-27b-3p in 5 OS cell lines(SAOS-2,143B,MG63,U20S,SW1353);(2)The SAOS-2 cell line was selected as the research object,and Western Blot technology was always used to detect the expression of SMAD1 in the three groups of Blank,NC and miR-27b-3p inhibitor groups;using dual luciferase reporting experiments The targeting relationship between miR-27b-3p and SMAD1 was determined.(3)The effects of miR-27b-3p on the proliferation,migration,invasion,and apoptosis of SAOS-2 cells were studied by MTT,Transwell migration experiments,Transwell invasion experiments,and flow cytometry.Results:1.Bioinformatics test results showed that miR-27b has a binding site with SMAD1-UTR;143B cells were used as a control group,and the 2~-??Ct??Ct method was used for relative quantitative analysis.The results showed that miR-27b-3p In the SAOS-2 cell line,the amount of reduction was the most fold,so SAOS-2 cells were selected for subsequent experiments.The results of Western Blot experiments showed that silencing miR-27b-3p can reduce the expression of SMAD1 in SAOS-2 cells.The results of the double luciferase report experiment showed that miR-27b-3p may regulate gene expression by binding to the3'UTR site of SMAD1.3.MTT cell proliferation experiments show that inhibiting miR-27b-3p can inhibit the proliferation of osteosarcoma cells.Transwell migration experiments show that inhibition of miR-27b-3p can inhibit the migration ability of osteosarcoma cells.Transwell invasion experiments showed that inhibition of miR-27b-3p can inhibit the invasion ability of osteosarcoma cells.Flow cytometry results showed that miR-27b-3p can induce osteosarcoma cell apoptosis.Conclusion:miR-27b-3p may promote the invasion,proliferation,migration,and apoptosis of oste osarcoma cells by regulating the expression of SMAD1.Our results suggest that miR-27b-3p may play an oncogene role in osteosarcoma cells.