Combinative Effects Of TRAM-34 And Carboplatin On Apoptosis And Cell Cycle Of Hela Cell Line

Objective: With the continuous increase in the incidence and mortality of cervical cancer,it is of great practical significance to explore a treatment plan with good curative effects and low side effects on cervical cancer.In this experiment,TRAM-34 and carboplatin were used in combination with HeLa cells for cervical cancer HeLa cells,and the effects of different dosing regimens on HeLa cell morphology,proliferation,apoptosis,and cell cycle were observed.To investigate whether the combination of TRAM-34 and carboplatin in HeLa cells is better than carboplatin alone in HeLa cells.Provides the possibility and relevant preliminary experimental evidence for the search for new chemotherapy-assisted drugs for clinically effective treatment of cervical cancer.Method:1.The cell proliferation inhibition rate of TRAM-34 and carboplatin was determined by CCK-8,and the half maximal inhibitory concentration of the drug was calculated according to the SPSS software.Finally,the next experiment was performed after screening the respective optimal concentrations.2.According to the determined IC50 group,the control group,TRAM-34 group,carboplatin group,and TRAM-34 combined carboplatin group were set.And then CCK-8 was used to determine the cell proliferation inhibition rate of HeLa cells at different time points,and use the corresponding statistical analysis.3.Using inverted phase contrast microscope to observe the changes of morphological changes at different time points(24h,48 h,72h)after cervical cancer HeLa cells were treated by diffrent dosing regimens.4.Annexin V-7AAD double staining was used to determine the apoptotic rate of HeLa cells treated with carboplatin and TRAM-34 alone and in combination at 24 h,48h and 72 h.5.The change of HeLa cell cycle was determined by PI staining.Carboplatin and TRAM-34 were used alone and in combination to change the cell cycle of HeLa cells at 24 h,48h,and 72 h.Result: 1.The results of CCK-8 analysis showed that the IC50 of carboplatin was 59.2 μg/ml and the IC50 of TRAM-34 was 46.5 μM.2.The results of CCK-8 assay showed that TRAM-34 and carboplatin alone or in combination with HeLa cells all inhibited cell proliferation,and the inhibitory effect increased with time(P<0.001).Carboplatin inhibited the proliferation of HeLa cells more strongly than TARM-34.When TRAM-34 was combined with carboplatin,the inhibitory effect of HeLa cells on proliferation was stronger than that of carboplatin alone,and the inhibitory effect was enhanced with time.3.Inverted phase contrast microscopy showed that: HeLa cells shrink,float,round,reduced cytoplasm,nuclei pyknosis,with the drug gradually extended the time after the morphological change is more obvious.4.The results of PE Annexin V-7AAD staining showed:Both TRAM-34 and carboplatin alone promoted apoptosis in HeLa cells(P<0.001),and with the extension of time to promote the role of enhanced.Carboplatin promoted the apoptosis of HeLa cells more strongly than TARM-34.The combination of TRAM-34 and carboplatin promotes apoptosis in HeLa cells more strongly than carboplatin alone,and with the increase of time to promote the role of enhanced.5.The results of PI staining revealed that When TRAM-34 and carboplatin were used alone or in combination with HeLa cells,they all had a blockade effect on cell cycle(P<0.001),and the promotion effect was enhanced with time.The blockade effect of carboplatin on HeLa cells mainly showed S phase arrest.The blockade effect of TARM-34 on HeLa cells was mainly G1 arrest.The combination of TRAM-34 and carboplatin in HeLa cells mainly showed G1+S phase arrest.Conclusion: 1.Both Carboplatin and TRAM-34 alone used in cervical cancer HeLa cells inhibited cell proliferation and promoted apoptosis,and was positively correlated with the time of action.The effect of carboplatin is better than TRAM-34;it indicates that both have anti-tumor effect;2.The combination of carboplatin and TRAM-34 in cervical cancer HeLa cells showed that the inhibition of cell proliferation and apoptosis was superior to that of carboplatin alone.It is suggested that carboplatin and TRAM-34 have a synergistic effect in inhibiting cell proliferation and promoting apoptosis of HeLa cells.It is speculated that TRAM-34 can enhance the chemotherapy effect of carboplatin in the treatment of cervical cancer;3.Carboplatin and TRAM-34 alone and in combination used in cervical cancer HeLa cells have a cell cycle arrest effect.Carboplatin mainly showed S phase arrest;TRAM-34 mainly showed G1 arrest;TRAM-34 combined with carboplatin mainly showed G1+S phase arrest,which was a superposition of G1 phase blockade effect of TARM-34 and S phase arrest effect of carboplatin.