The Role And Its Mechanism Of Twist On Cisplatin Resistance Of Cervical Cancer In Hypoxic Microenvironment

Objective:Investigate the role and its mechanism of Twist on cisplatin resistance of cervical cancer in hypoxia microenvironment,to provide an experimental basis to improve the therapeutic efficacy for cervical cancer.Methods:The expressions of HIF-1α, Twist, MDR1 in middle and advanced stage cervical cancer tissue microarray chip which contains 78 cases were evaluated by immunohistochemistry method to study their relationship with clinical pathological characteristics and the correlation among them; CoC12 were used to mimic hypoxia environment. Hela Cells were divided into two groups, normal oxygen group and hypoxia group. Hela cells were treated with different concentration cisplatin and different time. The cell inhibition rate was measured and the working concentration and treatment duration of Cisplatin was optimized by MTT method; The expressions of HIF-1α, Twist and MDR1 proteins of cells were measured by Immunofluorescence; The expressions of HIF-1α, Twist and MDR1 in normal oxygen group, cisplatin group, hypoxic group and hypoxic cisplatin group were determined by Western blot and the correlation among them.Results:(1)The expressions of HIF-1α, Twist and MDR1 in 78 cases of cervical cancer tissue were 69.2%, 71.8% and 85.6%. The expressions of HIF-la, Twist and MDR1 in cervical cancer were significantly related to tumor Lymphatic Metastasis,differentiation degree and FIGO stage respectively (P<0.05). The expressions of HIF-1α, Twist and MDR1 gene were gradually increased with Lymphatic Metastasis, tumor differentiation degree reducing and FIGO clinical stage and there was significantly positive correlation between them(P<0.05).(2)Hela cells proliferation were inhibited by cisplatin either in normal oxygen group and hypoxia group. The inhibiton rate increased with the concentration and work time increased (P<0.05). The optimized working concentration and work time of Cisplatin was 10-5mol/L and 24h The IC50 value were 10-52 mol/L and 10-4.5 mol/L of Cisplatin respectively in normal oxygen group and hypoxia group, there was significantly difference between them(P<0.05).(3)The expressions of HIF-1α, Twist and MDR1 of He la cells in hypoxia microenvironment were significantly higher than those in the normal oxygen cells group (P<0.05), and there was significantly positive correlation among them(P<0.05).(4)Compare to the normal oxygen group, The expressions HIF-a, Twist and MDR1 was significantly higher than those in other three groups and which were highest in hypoxic cisplatin group (P<0.05), there were significant positive correlations among them in hypoxic group and hypoxic cisplatin group (P<0.05).Conclusions:The hypoxia microenvironment may be related to the cisplatin resistance of cervical cancer.The possible mechanism is that the HIF-1α/Twist Pathway activated and MDR1 upregulated, then promote the occurrence of cervical cancer cisplatin resistance.