Abstract：Objective To explore the effect of total glycosides in Glaucescent Fissistigma root on glycolysis, apoptosis and autophagy of lung adenocarcinoma cells through Kirsten rats arcoma viral oncogene homolog (KRAS). Methods The NCI-H1299 cells were cultured in vitro and divided into the control group, the low-dose group of total glycosides in Glaucescent Fissistigma root, the high-dose group of total glycosides in Glaucescent Fissistigma root, the group of total glycosides in Glaucescent Fissistigma root+Vector, and the group of total glycosides in Glaucescent Fissistigma root+KRAS. The control group was treated with normal cultue medium, while the low-dose group received final concentration of 5 mg/ml of total glycosides in Glaucescent Fissistigma root, the high-dose group received final concentration of 20 mg/ml of total glycosides in Glaucescent Fissistigma root. The cells in the total glycosides in Glaucescent Fissistigma root+Vector group were transfected with a negative control vector first, and then the cells were treated with 20 mg/ml of total glycosides in Glaucescent Fissistigma root. After the cells were transfected with a KRAS overexpression vector, the total glycosides in Glaucescent Fissistigma root+KRAS group were treated with 20 mg/ml of total glycosides in Glaucescent Fissistigma root. The expression of KRAS mRNA in cells of each group was detected by qRT-PCR, and the expression of KRAS protein in cells of each group was detected by Western blot. And the cell proliferation inhibition rate was detected by MTT method. The glucose consumption and lactic acid content of cells in each group were detected by kit. The apoptosis rate was detected by flow cytometry, and the expression levels of apoptotic and autophagy-related proteins in cells of each group were determined by Western blot. Results The expression levels of KRAS, glucose consumption, lactic acid content, and Bcl-2 protein in the low-dose group and the high-dose group were lower than those in the control group, the cell proliferation inhibition rate and apoptosis rate, LC3-Ⅱ/Ⅰ, beclin-1 protein and Bax protein levels were higher than those in the control group, and the differences were statistically significant (P<0.05). The levels of KRAS, glucose consumption, lactic acid content, and Bcl-2 protein expression in the cells of the total glycosides in Glaucescent Fissistigma root+KRAS group were higher than those of the total glycosides in Glaucescent Fissistigma root+Vector group, and the differences were statistically significant (P<0.05). The levels of cell proliferation inhibition, apoptosis rate, LC3-Ⅱ/Ⅰ, Bax protein and beclin-1 protein in the the total glycosides in Glaucescent Fissistigma root+KRAS group were lower than those in the the total glycosides in Glaucescent Fissistigma root+Vector group, and the differences were statistically significant (P<0.05). There were no significant differences in the various indexes between the the high-dose group of total glycosides in Glaucescent Fissistigma root and the total glycosides in Glaucescent Fissistigma root+Vector group (P>0.05). Conclusion The total glycosides in Glaucescent Fissistigma root can inhibit the expression of KRAS, inhibit the glycolysis process of lung adenocarcinoma cells, and promote the apoptosis and autophagy of lung adenocarcinoma cells, thereby inhibiting the lung adenocarcinoma cells.
京公网安备 11010502046607号