Mechanism Study on the Effect of Gegen Qinlian Decoction on the Occurrence and Development of Colorectal Cancer by Regulating the lncRNA-TUSC7/miR-182/TBX5 Signal Axis
Objective:To explore the mechanism of Gegen Qinlian Decoction on the occurrence and develop-ment of colorectal cancer(CRC)by regulating the lncRNA-TUSC7/miR-182/TBX5 signaling axis.Methods:1)CRC cells HCT-116 were cultured in vitro,HCT-116 cells were divided into the control group,oe-NC group,oe-TUSC7 group,inhibitor-NC group,miR-182 inhibitor group,oe-TUSC7+miR-NC group,and oe-TUSC7+miR-182 mimic group.RT qPCR was applied to detect the expression of lncRNA-TUSC7,miR-182,and TBX5 mRNA;CCK-8 was applied to detect cell proliferation;flow cytometry was applied to detect cell apoptosis rate;Western blotting was applied to detect the expression levels of TBX5,Ki67 and Caspase-9 proteins in cells.Double Lucifer-ase reporter gene experiment was applied to detect the targeting relationship between miR-182 and lncRNA-TUSC7 and TBX5.2)The rat CRC model was established by subcutaneous injection of DMH,and was divided into the model group,Gegen Qinlian Decoction group,Gegen Qinlian Decoction+LV-NC group,and Gegen Qinlian De-coction+LV-shTUSC7 group;another normal group(no treatment)was set up.The number,mass,and volume of tumors were measured;RT-PCR was applied to detect the expression of TUSC7,miR-182,and TBX5 mRNA in tu-mor tissue;Western blotting was applied to detect the expression of TBX5,Ki67,and Caspase-9.Results:1)Com-pared with the control group and oe-NC group,the expression of TUSC7,TBX5 mRNA,apoptosis rate,the expres-sion of TBX5,and Caspase-9 proteins in cells in the oe-TUSC7 group were obviously increased,the expression of miR-182,OD450 value(24,48 hours),and the expression of Ki67 were obviously reduced(P＜0.05);compared with the inhibitor-NC group,the expression of TBX5 mRNA,apoptosis rate,the expression of TBX5,and Caspase-9 proteins in cells in the miR-182 inhibitor group were obviously increased,the expression of miR-182,OD450 value(24,48 hours),and the expression of Ki67 were obviously reduced(P＜0.05);compared with the oe-TUSC7+miR-NC group,the expression of TBX5 mRNA,apoptosis rate,the expression of TBX5,and Caspase-9 proteins in cells in the oe-TUSC7+miR-182 mimic group were obviously reduced,the expression of miR-182,OD450 value(24,48 hours),and the expression of Ki67 were obviously increased(P＜0.05);the double Luciferase reporter gene experiment verified that miR-182 had a targeted regulatory relationship with TUSC7 and TBX5.2)The in vi-vo experimental results showed that compared with the normal group,the number of tumors,tumor mass,tumor vol-ume,the expression of miR-182,and the expression of Ki67 protein in the model group were obviously increased,the expression of TUSC7 and TBX5 mRNA,and the expression of TBX5 and Caspase-9 proteins in tumor tissue were obviously reduced(P＜0.05);compared with the model group,the number of tumors,tumor mass,tumor vol-ume,the expression of miR-182,and the expression of Ki67 protein in the Gegen Qinlian Decoction group were ob-viously reduced,the expression of TUSC7 and TBX5 mRNA,and the expression of TBX5 and Caspase-9 proteins in tumor tissue were obviously increased(P＜0.05);compared with the Gegen Qinlian Decoction group and the Ge-gen Qinlian Decoction+LV-NC group,the number of tumors,tumor mass,tumor volume,the expression of miR-182,and the expression of Ki67 protein in the Gegen Qinlian Decoction+LV-shTUSC7 group were obviously in-creased,the expression of TUSC7 and TBX5 mRNA,and the expression of TBX5 and Caspase-9 proteins in tumor tissue were obviously reduced(P＜0.05).Conclusion:Gegen Qinlian Decoction may up-regulate TUSC7 and sponge miR-182,thereby promoting TBX5 expression and inhibiting the growth of CRC cells.
Colorectal cancerGegen Qinlian DecoctionlncRNA-TUSC7/miR-182/TBX5 signal axisRats
NETL
NSTL
万方数据
