Loquat Leaf Extract Regulates TGFBR3, Affects PI3K/Akt Signaling Pathway And Inhibits Metastasis Of Lung Cancer Cell

Objective: To investigate the effect of loquat leaf extract in regulating TGFBR3 on lung cancer invasion and metastasis,and to investigate whether it is associated with tumor-associated PI3K/Akt signaling pathway at the cellular level initially.Methods:1.By analyzing clinical samples,we analyzed the difference of TGFBR3 protein expression in lung adenocarcinoma tissues and their adjacent paraneoplastic tissues by immunohistochemical assay,and analyzed the correlation between TGFBR3 expression and clinical stage and lymphatic metastasis of lung adenocarcinoma cell carcinoma patients.2.Two lung cancer cell lines,A549 and NCI-H292,were cultured,and A549 cells and NCI-H292 cells were treated with loquat leaf extract,supplemented with blank control,negative control of DMSO group and cisplatin as positive control,and cell crawl and histochemical staining were used to observe whether loquat leaf extract affected the expression level of TGFBR3 in lung cancer cells.3.The expression of TGFBR3 in A549 cells and NCI-H292 cells was detected by Western Blot technique,and the overexpression plasmid vector and interference plasmid vector were constructed respectively.The cells were divided into four groups: TGFBR3 overexpression group,TGFBR3 overexpression negative control group,TGFBR3 interference group,TGFBR3 interference negative control,and four groups of blank control group,DMSO negative control group,loquat leaf extract experimental group and cisplatin positive control group were set up in each major group,totaling 16 groups of cells.These 16 groups were subjected to functional experimental studies of cell scratching,cellular vesicles and other migration and invasion,and the effects of loquat leaf extract and TGFBR3 on the invasive metastatic ability of lung cancer cells were analyzed.4.The expression levels of PI3 K,p-PI3 K,AKT,and p-Akt proteins in the above 16 groups of cells were detected at the cellular level by Western Blot assay,and it was determined that loquat leaf extract regulated TGFBR3 to influence PI3K/Akt signaling pathway to inhibit tumor cell metastasis.Results:1.In the tissue sections of 41 patients with adenocarcinoma of the lung,the expression of TGFBR3 protein was significantly lower in liver cancer tissues than in adjacent paracancerous tissues,and the expression of TGFBR3 protein correlated with the clinical stage and lymphatic metastasis of lung cancer.The results of cell crawl confirmed that the loquat leaf extract group could affect the TGFBR3 protein expression level in lung cancer cells compared with the blank control group and DMSO-negative control group.2.TGFBR3 showed low expression level in A549 cell line and high expression level in H292 cell line.After performing overexpression and interference plasmid constructs,the results of 16 groups of cell scratch and transwell assays showed that each of the four major groups had reduced migratory invasive ability of cells compared to the interference group in the overexpression group,and both the loquat leaf extract and cisplatin groups within the four groups inhibited the migration and metastatic ability of cells.3.Western Blot assay showed that the expression levels of p-PI3 K and p-Akt proteins were down-regulated after TGFBR3 overexpression group in four major groups of cells,p-PI3 K and p-Akt protein levels were also down-regulated to different degrees in the loquat leaf extract group and cisplatin-positive control group within the four groups.Conclusion:1.Loquat leaf extract up-regulated TGFBR3 protein expression in lung cancer.2.Loquat leaf extract regulates TGFBR3 to inhibit the migration and invasion ability of lung cancer cells.3.Loquat leaf extract regulates TGFBR3 to inhibit the expression of p-PI3 K and p-Akt,which can partially restore the function of metastasis inhibition in TGFBR3 low expression lung cancer cells.