Effects Of 1,25?OH?₂D₃ Combined With Cisplatin On Proliferation And Expression Of P27 Protein In Cervical Cancer Hela Cells

Objective: Cervical cancer is one of the most serious diseases that threaten women's health now.China's new cases of cervical cancer each year are 131,000,and 53,000 people die each year from the disease.Over the past 30 years,due to the popularity of cervical cancer screening,worldwide cervical squamous cell carcinoma morbidity and mortality were significantly decreased,but the incidence of adenocarcinoma is an upward trend,and epidemiological investigation also shows that in recent years,the incidence of cervical cancer in China shows a younger trend.Changes in the incidence of patients and pathological types,and changes in the choice of treatment mode make cervical cancer chemotherapy attract more and more attention.But chemotherapy drugs were restricted in clinical,because of drug resistance and side effects.So looking for a new type of tumor treatment drugs or chemotherapy sensitization drugs,is a hot topic of cancer treatment.In this study,Hela cells in vitro were treated with 1,25-dihydroxyvitamin D3 [1,25?OH?₂D₃] and cisplatin?DDP?to observe the influence on proliferation in cervical cancer.The aim of this study is to provide experimental evidence for the anti-tumor effect of 1,25?OH?₂D₃ and improving the curative effect of clinical cervical cancer.Methods: Cervical cancer Hela cells in logarithmic growth period were tested with 1,25?OH?₂D₃ and cisplatin.MTT assay was used to detect the two drugs on the proliferation of Hela cells.The cell cycle distribution of Hela cells were detected by flow cytometry.Western blotting was used to detect the expression of P27 protein in cervical cancer Hela cells.Results: 1.MTT assay showed that 1,25?OH?₂D₃ in the range of 10-8¹0-6 mol/L had no significant relationship with the proliferation of cervical cancer Hela cells?P>0.05?,but was closely related to the time of action?P<0.05?,and the inhibitory effect of 1,25?OH?₂D₃ monotherapy at 10-6mol/L on the proliferation of Hela cells was the most significant after 72 h.Cisplatin concentration in the range of 5²0?g/ml of cervical cancer Hela cells proliferation was significantly inhibited,showing a Concentration-time-dependent?P<0.01?.The maximum inhibition rate was 96.02±0.24% at the concentration of 20?g/ml for 72 h.The inhibitory effect of 1,25?OH?₂D₃ combined with DDP or 1,25?OH?₂D₃ pretreatment after 12 h sequential DDP on the proliferation of Hela cells was significantly higher than that of other groups?P<0.01?.1,25?OH?₂D₃ and DDP different combinations of different ways to cervical cancer Hela cell proliferation inhibition was statistically significant?P<0.01?.The proliferation inhibition effect to the maximum when 1,25?OH?₂D₃ pretreatment after 12 h sequential DDP on cervical cancer Hela cells.Q values are equal to 1.76 and 2.56,respectively>1.15,suggesting that 1,25?OH?₂D₃ and DDP act on cervical cancer Hela cells with synergistic anti-tumor activity,which is calculated by Jinzhengjun Q method?formula:q=EA+B/EA+EB-EA×EB?.2.Flow Cytometry was used to detect the cell cycle distribution of cervical cancer Hela cells after 48 h of treatment.The percentage of G0/G1 phase in the cell cycle of group A,B,C,D and E was 58.31±0.06%,59.85±0.67%,66.00±0.19%,72.15±0.39%,75.94±0.27%.Compared with the control group,the proportion of G0/G1 phase in group B,C and D was increased,with significant statistical difference?P<0.01?.And there was significant difference between group D and group C?P<0.01?.3.Western Blot analysis showed that 1,25?OH?₂D₃ and DPP treated with cervical cancer Hela cells 48 h,the expression of P27 protein?expressed in grayscale values?of each group was 0.12±0.01,0.25±0.01,0.56±0.01,0.77±0.02,0.88±0.02.Compared with the control group,the expression levels of P27 protein in group A,B,C and D were significantly higher?P<0.01?.And the expression of protein in D group was the most significant?P<0.01?.Conclusion:1.Cisplatin inhibits proliferation of cervical cancer Hela cells in a concentration-time-dependent manner.1,25?OH?₂D₃ inhibited the proliferation of cervical cancer Hela cells only showed a certain time-dependent.2.1,25?OH?₂D₃ can significantly increase the inhibitory effect of cisplatin on proliferation of cervical cancer Hela cells.The inhibitory effect was more significant When 1,25?OH?₂D₃ pretreatment.3.1,25?OH?₂D₃ combined with cisplatin can induce cervical cancer Hela cell cycle arresting in G0/G1 phase.4.1,25?OH?₂D₃ combined with cisplatin can up-regulate the expression of P27 protein in Hela cells of cervical cancer and induce apoptosis,and then play a synergistic anti-tumor effect.