Experimental Research For Relationship Between MTSS1 And Cervical Cancer

BackgroundsCervical cancer is one of common malignancy of Gynecology, a serious threat to women’s health, and the incidence increased in recent years, despite early detection combines radical surgery can improve the prognosis of patients with cervical cancer, there are still some patients with the emergence of local recurrence or distant metastasis, thus it is particularly urgent need to study the molecular mechanism of cervical cancer incidence to guide clinical diagnosis and treatment. Studies have reported that MTSS1 (Metastasis suppressor 1) associated with metastasis and disease progression of a variety of malignant tumors, it may provide guidance for the research of molecular mechanismsin cervical cancer, development and application of new drugs in clinical treatment to study the expression of MTSS1 in cervical cancer progression.ObjectiveTo detect the expression of MTSS1 in cervical tissues and clarify its relationship to cervical cancer; To explore the correlation between MTSS1 and Shh signaling pathway in cervical cancer; To investigate the effect of Cisplatin (DDP) on MTSS1、ERK and AKT signaling pathways in Hela cell and to explore the molecular mechanism of this gene in cervical cancer.Methods1. q-PCR and Western blot were used to detect the expression of MTSS1 in different cervical cancer tissue.2. q-PCR was used to detect the mRNA expression level of MTSS1 and Shh signaling pathway in different cervical cancer tissue; Western blot and IHC (immunohistochemistry) were used to detect the protein level of MTSS1 and Glil.3. In vitro cultured Hela cells were treated with gradient concentration of DDP for 24 h and 48 h, CCK-8 assay was used to determine the cytotoxicity; q-PCR was used to test the expression of MTSS 1 after treated with DDP; Moreover, Western blot was used to determine the protein level of MTSS1, p-ERK and p-AKT.Resultsl.The expression of MTSS 1 in middle-late stage of cervical cancer tissue was significantly higher than that of normal tissue or early stage through q-PCR, there are significantly statistical difference among the three groups (P<0.01); Western blot results showed that the expression of MTSS 1 was poorly correlated with age, tumor differentiation and lymphnode metastasis in cervical cancer tissue (P>0.05), but related to the clinical stages (P<0.01).2. q-PCR results showed that the expression of MTSS 1, Ptch, Smo and Glil increased gradually with the clinical stage progressing, All the four genes of the middle-late stage cervical cancer group were the highest (P<0.01); Western blot showed that MTSS1 protein expression in middle-late stage of cervical cancer tissue was significantly higher than that of early stage cervical cancer group or normal cervical group (P <0.05); IHC results showed that MTSS1 and Glil protein expression in cervical cancer tissue group were higher than that of normal cervical tissue group.3. Treated with DDP for 24 or 48 h, the proliferation of Hela cells was inhibited significantly (P<0.05), the value of cells half inhibitory concentration (IC50) were 4.14 μumol·L-1 and 11.82 μmol·L-1; q-PCR results showed that MTSS1 mRNA expression was DDP concentration-dependently (correlation coefficient r24h=-0.965, P<0.01;r48h=-0.953, P<0.01); Western blot showed that the protein levels of MTSS 1, p-ERK and p-AKT expression were declined significantly with DDP concentrations increasing.Conclusion1. The expression of MTSSl is related to the stage of cervical cancer, and MTSSl may play an important role in cervical cancer.2. MTSS1 expression level in cervical cancer is consistent with the Shh signaling pathway activation, with the progressing of cervical cancer, both MTSS1 and Shh signaling activation got enhanced, suggesting a positive correlation of MTSS1 and Shh pathway as potential therapeutic targets in cervical cancer.3. DDP treatment inhibits Hela proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.