Mechanism of regulation of miR-433-3p/SRC by baicalin combined with oxaliplatin on proliferation and invasion of gastric cancer cells
Objective To investigate the effect of baicalin combined with oxaliplatin to regulate miR-433-3p/SRC on the proliferation and invasion of gastric cancer cells.Methods SGC-7901 cells were divided into control group,baicalein(100,200,and 300 μmol·L-1)group,oxaliplatin group(33 μmol·L-1),and combination therapy group(300 μmol·L-1 baicalein and 33 μmol·L-1 oxaliplatin).They were divided into miR-433-3p group,miR-NC group,anti-miR-433-3p group,anti-miR-NC group,pcDNA-SRC group,si-SRC group,miR-433-3p+combination therapy group,anti-miR-433-3p+combination therapy group,pcDNA-SRC+combination therapy group, si-SRC + combination therapy group, and miR-433-3p + pcDNA-SRC + combination therapy group. CCK-8 method was used to detect cell proliferation ability, transwell assay was used to detect cell invasion ability, detection of cell apoptosis rate using flow cytometry, real time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of miR-433-3p in cells,double luciferase assay was used to detect the targeting relationship between miR-433-3p and SRC, and Western blotting was used to detect the expression of SRC protein in cells. Results Compared with control group, the cell proliferation inhibition rate, apoptosis rate and miR-433-3p expression increased and the cell invasion number and SRC protein expression decreased in baicalin (100,200, and 300 μmol·L-1), oxaliplatin and combination therapy groups (P ＜ 0.05). Compared with the oxaliplatin group and baicalin 300 μmol·L-1 group, the combination therapy group showed a significant increase in cell proliferation inhibition rate and apoptosis rate, while the number of cell invasions decreased significantly. Compared with baicalin 300 μmol·L-1 group, the expression of miR-433-3p significantly increased in combination therapy group (P ＜ 0.05) Compared with the oxaliplatin group, the expression of SRC protein was significantly reduced in combination therapy group (P ＜ 0.05). Compared with control group,miR-433-3p expression was elevated and SRC protein expression was decreased in cells of miR-433-3p group, and miR-433-3p expression was decreased and SRC protein expression was increased in cells of anti-miR-433-3p group (P ＜ 0.05). The cell proliferation inhibition rate and apoptosis rate of the miR-433-3p + combination therapy group were significantly higher than those of the combination therapy group, and the number of cell invasions was significantly lower than that of the combination therapy group (P ＜ 0.05). The cell proliferation inhibition rate and apoptosis rate of the anti miR-433-3p+combination therapy group were significantly lower than those of the combination therapy group, and the number of cell invasions was significantly higher than that of the combination therapy group (P ＜ 0.05). The luciferase activity of SRC-WT in the miR-433-3p group was significantly lower than that in the miR-NC group (P ＜ 0.05). The expression of SRC protein in si-SRC group cells was significantly lower than that in the control group (P ＜ 0.05), while the expression of SRC protein in pcDNA-SRC group cells was significantly higher than that in the control group (P ＜ 0.05). Compared with the pcDNA-SRC group, the expression of SRC protein in the miR-433-3p+pcDNA-SRC group cells was significantly reduced (P ＜ 0.05). The cell proliferation inhibition rate and apoptosis rate of the si-SRC+combination therapy group were significantly higher than those of the combination therapy group, and the number of cell invasions was significantly lower than that of the combination therapy group (P ＜ 0.05). The cell proliferation inhibition rate and apoptosis rate of the pcDNA-SRC+combination therapy group were significantly lower than those of the combination therapy group, and the number of cell invasions was significantly higher than that of the combination therapy group (P ＜ 0.05). Compared with the pcDNA-SRC+combination group, the miR-433-3p+pcDNA-SRC+combination group showed a significant decrease in cell proliferation inhibition rate and apoptosis rate (P ＜ 0.05), and a significant increase in cell invasion number (P ＜ 0.05). Conclusion Baicalin combined with oxaliplatin can inhibit the proliferation and invasion of gastric cancer cells through miR-433-3p-targeted regulation of SRC and induce apoptosis of gastric cancer cells.
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