Study On The Mechanism Of Huisheng Oral Liquid Regulating EMT Of Lung Cancer Cells Via MiR-200b/ZEB-1 Signal Loop

ObjectiveTo study the mechanism of Huisheng Oral Liquid through the miR-200 b / ZEB-1signal loop to regulate the epithelial-mesenchymal transition(EMT)of lung cancer to promote tumor recovery Development provides experimental basis.Methods1.In vitro culture of Lewis lung cancer cells,after intervention by Huisheng oral liquid,cisplatin,Huisheng oral liquid + cisplatin-containing serum,using flow cytometry to observe the cycle change of Lewis lung cancer cells.2.56 healthy C57 BL / 6J mice were divided into 4 groups according to th e random number table,14 in each group,which were model group,cisplatin group,Huisheng oral liquid group,combined medication group(huisheng oral l iquid + cisplatin),in Lewis lung cancer cell suspension was injected subcutane ously in the right armpit of mice to establish Lewis mouse lung cancer model.After successful modeling,the experimental mice in each group were interven ed.The Huisheng oral liquid group was given intragastric administration of Hu isheng oral solution,the cisplatin group was given intraperitoneal injection of c isplatin,and the combined treatment group was given intragastric administration of Huisheng oral solution+intraperitoneal injection of cisplatin.The model gr oup was given intragastric saline.After 2 weeks of intervention,the mice were sacrificed by dislocation of the cervical spine,and the tumor was dissected to observe the ultrastructure of lung cancer cells under a transmission electron mi croscope.3.RT-PCR technology was used to detect the expression of miR-200 b gene in lung cancer cells,and Western Blot was used to detect the expression of ZEB-1,E-cadherin and Vimentin protein in lung cancer cells.Results1.The effect of Huisheng oral liquid on the cell cycle of Lewis lung cancerCompared with the control group,the proportion of Lewis lung cancer cells in the G1 phase of the Huisheng oral liquid group,cisplatin group,and combination medication group was significantly increased,while the proportion of cells in the S and G2 phases was significantly reduced(P<0.05).Comparing the three treatment groups of Huisheng oral liquid group,cisplatin group and combination therapy group,the proportion of Lewis lung cancer cells in G1 phase increased and the proportion of S phase decreased in the following order: cisplatin group> combination group>huisheng oral liquid Group(P<0.05).2.The effect of Huisheng oral liquid on the ultrastructure of Lewis lung cancer cellsThe lung cancer cells in the model group had a complete structure and were tightly connected to the surrounding cells.The chromatin in the cells wa s uniform,the proportion of nucleoplasm was increased,the degree of nuclear atypia was high,and there was no swelling or damage to the organelles.Hu isheng oral liquid group: The structure of lung cancer cells is still clear,the cell volume is significantly reduced,the nucleus is irregular,the chromatin in the nucleus is fragmented,and the edge is distributed.It is distributed inside the nuclear membrane,and mitochondrial vacuole-like degeneration is seen in the cytoplasm.Cisplatin group: The structure of lung cancer cells is still cle ar,the nucleus is irregular,the chromatin in the nucleus is concentrated,and mitochondrial vacuole-like degeneration and apoptotic bodies are visible.Comb ined medication group: Apoptosis of lung cancer cells is more common,the c ell structure is incomplete,the chromatin in the nucleus is fragmented,and it is densely distributed,showing mitochondrial vacuole-like degeneration and auto phagosome formation.3.The effect of Huisheng oral liquid on the expression of miR-200 b in lung cancer cellsCompared with the model group,the miR-200 b expression in the Huisheng oral liquid group and the combination medication group was significantly increased(P<0.05),and there was no statistical difference between the cisplatin group and themodel group(P>0.05);the Huisheng oral liquid group and the cisplatin group Compared with the combination group and the combination group,the miR-200 b expression in the Huisheng oral liquid group was significantly higher than that in the combination group and the cisplatin group(P<0.05).The miR-200 b expression in the combination medication group was significantly higher than that in the cisplatin group(P<0.05).The expression of miR-200 b from high to low is as follows:Huisheng oral liquid group>combination drug group>cisplatin group(P<0.05).4.The effect of Huisheng oral liquid on the expression of ZEB-1 protein,E-cadherin protein and Vimentin protein in lung cancer cells(1)ZEB-1 protein expression: Compared with the model group,the ZEB-1expression in the Huisheng oral liquid group,the cisplatin group and the combination medication group was significantly reduced(P<0.05);compared between the treatment groups,the ZEB-1 expression was increased by In descending order: Huisheng oral liquid group>cisplatin group>combination medication group(P<0.05).(2)E-cadherin protein expression: Compared with the model group,the E-cadherin expression in the Huisheng oral liquid group,the cisplatin group,and the combination medication group was significantly increased(P<0.05);compared between the treatment groups,the E-cadherin expression was determined by The order from high to low is: combination medicine group> cisplatin group / huisheng oral liquid group(P<0.05).(3)Vimentin protein expression: Compared with the model group,the expression of Vimentin in the Huisheng oral liquid group,the cisplatin group and the combination medication group was significantly reduced(P<0.05);compared between the treatment groups,the expression of Vimentin from high to low was:Liquid group>cisplatin group>combination medication group(P<0.05).Conclusion1.Huisheng oral liquid can induce Lewis lung cancer cell cycle to stagnate in G1 phase,and obviously inhibit the growth of Lewis lung cancer cells.2.The combination medication group has better effect on inhibiting the EMTprocess of lung cancer cells than Huisheng oral liquid or cisplatin alone.3.Huisheng oral liquid can inhibit the EMT of lung cancer cells through miR-200 b / ZEB-1 signal loop,which may be one of the mechanisms of Huisheng oral liquid in treating lung cancer.