Molecular Mechanism Of Lactate And Traditional Chinese Medicine On The Proliferation Of Non-small Cell Lung Cancer

Objective: This study aims to explore the correlation between the expression of lactate and GPR81 and its possible molecular mechanism in human non-small cell lung cancer cell lines,and provide a new theoretical basis for the treatment of clinical tumors.Methods: Human non-small cell lung cancer cell lines A549 and H1299 were selected as research subjects.(1)At the protein level,the STAT3 activity and the expression of GPR81 protein were detected by Western blot;(2)at the transcriptional level,the GPR81 promoter activity was detected by the dual luciferase reporter gene method;(3)Transfection of STAT3 cDNA or STAT3 siRNA and stimulated with different concentrations of lactate to detect the expression of GPR81 protein in A549,H1299 cells;(4)STAT3 cDNA,GPR81 promoter with Renilla luciferase rgsorfer gene were cotransfected in A549 and H1299 cells,and the effect of STAT3 cDNA on GPR81 promoter activity was detected by dual luciferase reporter gene method.Result: In A549 and H1299 cells:(1)STAT3 activity increased with increasing lactate concentration,GPR81 protein expression was up-regulated with increasing lactate concentration;(2)GPR81 promoter activity increased with increasing lactate concentration;(3)After transfection of STAT3 cDNA or STAT3 siRNA,p-STAT3 protein expression is consistent with GPR81 protein expression;(4)STAT3 cDNA can enhance GPR81 promoter activity.Conclusion: 1.Lactate can activate the phosphorylation activation of STAT3 and the expression of GPR81 protein in A549 and H1299 cells in a concentration-dependent manner.2.STAT3 acts as a transcription factor to promote the expression of GPR81 at the transcriptional level.Objective: To study the effect of Zhongjiefeng(ZJF)on the proliferation of human nonsmall cell lung cancer A549 and H1299,and the specific molecular mechanism.Methods: Firstly,MTT was used to study the inhibitory effect of different concentrations of ZJF on the growth of A549 and H1299 cells,and the cell proliferation inhibition rate was calculated.Flow cytometry(FCM)was used to determine the effects of ZJF on the cell cycle of A549 and H1299.The protein expressions of p21 and p-Smad3 in A549 and H1299 cells and TGF-?(SIS3 HC)inhibitors were detected by Western blot.Results: The proliferation inhibition rates of A549 and H1299 cells were all increased with the increase of the ZJF concentration.After 24 h of ZJF-induced cells,the proportion of G0/G1 phase cells was increased,indicating that ZJF promoted G0/G1 phase arrest of cells.Western blot results showed that the expression of p21 could be increased by upregulating the expression of p-Smad3.Conclusion: ZJF induces the expression of p21,a cyclin-kinase inhibitor,through the TGF-? signaling pathway,which blocks the cell cycle in the G0/G1 phase,thereby inhibiting the proliferation of lung cancer cells.