Mechanism Of Qige Powder On The Sensitivity Of Cisplatin To EC9706 Co-culture With HUVEC

ObjectiveTo study the effect of Qige Powder and cisplatin on the proliferation,apoptosis,cycle,miR-21 and downstream gene expression of EC9706 under co-culture of HUVEC,and to elucidate the mechanism of Qige Powder and cisplatin on esophageal cancer.MethodsRat serum containing medicine and control serum preparation;Screening the appropriate proportion of HUVEC and EC9706 in the indirect co-culture system,MTT assay was used to compare the difference between the effect of cisplatin on EC9706 and EC9706 in co-culture with HUVEC and EC9706,drug-containing serum percentage screening of inhibition on EC9706 in the HUVEC and EC9706 co-culture system,screening of the fixed ratio of cisplatin to serum containing.Effect of drug-containing serum and cisplatin on EC9706 under co-culture of HUVEC and EC9706 by factorial design.High and low expression of miR-21 lentivirus-mediated EC9706 response to cisplatin.Results1EC9706 proliferation in the HUVEC:EC9706?0:5?group was less than EC9706in any other co-culture group,The EC9706 proliferation in the HUVEC:EC9706?2:5?group were higher than that in HUVEC:EC9706?0:5?group and the other co-culture groups.The indirect co-culture system of HUVEC and EC9706 was performed by using HUVEC:EC9706 as 1:3?from 2:5?.2The IC₅₀ of EC9706 under co-culture condition to cisplatin is 0.878?g/m L.The IC₅₀ of EC9706 alone to cisplatin is 0.493?g/m L.The two are statistically significant?P<0.05?.3In the 2.5%serum group,the OD value of AD and BD decreased significantly compared with that of A?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically significant?P<0.05?.In the 5%serum group,the OD value of AD,B and BD decreased compared with that of A,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically significant?P<0.05?.In the 10%serum group,the OD value of AD,B and BD decreased compared with that of A,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically?P<0.05?.In the 20%serum group,the OD value of AD and BD decreased compared with that of A,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically significant?P<0.05?.4In the 1 group,the OD value of AD,B and BD decreased compared with that of A,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically significant?P<0.05?.In the two groups,the OD value of AD,B and BD decreased significantly compared with that of A?P<0.05?.Compared with B,the OD value of AD and BD decreased,the difference was statistically significant?P<0.05?.In the three groups,the OD value of AD and BD decreased,and the OD value of B increased,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,Statistically significant?P<0.05?.In the 4 groups,the OD value of AD and BD decreased,and the OD value of B increased,the difference was statistically significant?P<0.05?.Compared with B,the OD value of AD and BD decreased,Statistically significant?P<0.05?.5In the first group,the percentage of G1 and G2 in 1AD and 1BD decreased compared with 1A,and the percentage of G2 phase of 1B and 1BD increased,and the percentage of S phase of 1AD and 1BD increased,the difference was statistically significant?P<0.05?.Compared with 1B,the percentage of G1 phase of 1AD and 1BD decreased,the percentage of G2 phase of 1AD decreased,the percentage of G2 phase of1BD increased,and the S phase percentage of 1AD and 1BD increased,the difference was statistically significant?P<0.05?,the percentage of G1 phase of 1BD decreased,and the percentage of G2 phase of 1BD increased,the difference was statistically significant?P<0.05?.In the second group,the percentage of G1 phase in 2BD was statistically significant?P<0.05?compared with 2A.6In the first group,compared with 1A,the early apoptotic ratio of 1AD,1BD increased,the early apoptosis rate of 1B decreased,the late apoptotic ratio of 1AD and1BD increased,and the late apoptotic ratio of 1B decreased?P<0.05?.Compared with1B,the apoptotic ratio of 1D and 1BD was significantly higher than that of 1B?P<0.05?.Compared with 1AD,the apoptosis rate of 1BD?P<0.05?.The difference was statistically significant?P<0.05?.In the second group,the early and late apoptotic ratio of 2AD and 2BD increased compared with 2A,and the apoptotic ratio of 2B was lower than that of 2A?P<0.05?.Compared with 2B,2AD,2BD?P<0.05?.The difference was statistically significant?P<0.05?.Compared with 2AD,the late apoptotic ratio of 2BD increased,the difference was statistically significant?P<0.05?.7In the first group,the expression of miR-21 was decreased in 1AD,1B and 1BD compared with 1A,and the expression of PDCD4 in 1AD and 1BD increased,and the expression of Fas in 1BD increased,the difference was statistically significant?P<0.05?.Compared with 1B,the expression of miR-21 in 1AD and 1BD decreased,and the expression of PTEN in 1AD and 1BD increased,the difference was statistically significant?P<0.05?.In group 2,the expression of miR-21 in 2B increased,the expression of PDCD4 in 2AD and 2BD increased,the expression of PTEN in 2AD and2BD increased,the expression of Fas in 2AD and 2BD increased,and the expression of Fas in 2B decreased Was statistically significant?P<0.05?.Compared with 2B,the expression of miR-21 in 2AD increased,the expression of miR-21 in 2BD decreased,the expression of PDCD4 in 2AD and 2BD increased,the expression of PTEN in 2AD and 2BD increased,and the expression of Fas in 2AD and 2BD increased,the difference was statistically significant?P<0.05?.Compared with 2AD,the expression of miR-21 in2BD decreased,and the expression of Fas in 2BD increased,the difference was statistically significant?P<0.05?.8There has no significant difference in protein selected expression between the two groups.9Compared with control,the IC50 of miR-21 LV group increased,the difference was statistically significant?P<0.05?.Compared with miR-21-Inhibitor LV group,the IC50 of cisplatin in miR-21 LV group was significantly different?P<0.05?.Conclusion1HUVEC can promote the proliferation rate of EC9706 or inhibit its apoptosis.Co-culture of HUVEC and EC9706 attenuated the cytotoxicity of cisplatin to EC9706.2The serum concentration of Qige Powder increase the sensitivity of cisplatin of EC9706 co-cultured with HUVEC may be related to the increase of cell ratio in S phase and the down-regulation of miR-21 expression and up-regulation of Fas expression when collaborate with cisplatin.3MiR-21 plays a role in promoting EC9706 cisplatin resistance.