目的 探究黄芩苷联合奥沙利铂调控miR-433-3p/SRC对胃癌细胞增殖和侵袭的影响。方法 将SGC-7901细胞分为对照组、黄芩苷（100、200、300 μmol·L^-1）组、奥沙利铂（33 μmol·L^-1）组，联合用药（300 μmol·L^-1黄芩苷+33 μmol·L^-1奥沙利铂）组，miR-433-3p组、miR-NC组、anti-miR-433-3p组、anti-miR-NC组、pcDNA-SRC组、si-SRC组，miR-433-3p+联合用药组、anti-miR-433-3p+联合用药组、pcDNA-SRC+联合用药组、si-SRC+联合用药组、miR-433-3p+pcDNASRC+联合用药组。CCK-8检测细胞增殖能力；Transwell小室法检测细胞侵袭能力；流式细胞仪检测细胞凋亡率；实时荧光定量PCR （qRT-PCR）检测细胞中miR-433-3p表达；双荧光素酶报告检测miR-433-3p和SRC的靶向关系；Westernblotting检测细胞中SRC蛋白表达。结果 与对照组相比，黄芩苷（100、200、300 μmol·L^-1）组、奥沙利铂组、联合用药组细胞增殖抑制率、凋亡率和miR-433-3p表达显著增加（P<0.05），细胞侵袭数目、SRC蛋白表达显著减少（P<0.05）；与黄芩苷300 μmol·L^-1组、奥沙利铂组相比，联合用药组细胞增殖抑制率、凋亡率显著增加，细胞侵袭数目显著减少；与黄芩苷300 μmol·L^-1组比较，miR-433-3p表达显著增加（P<0.05）；与奥沙利铂组比较，SRC蛋白表达显著减少（P<0.05）。与对照组比较，miR-433-3p组细胞中miR-433-3p表达显著升高，SRC蛋白表达显著降低，anti-miR-433-3p组细胞中miR-433-3p表达显著降低，SRC蛋白表达显著升高（P<0.05）；miR-433-3p+联合用药组细胞增殖抑制率和凋亡率显著高于联合用药组，细胞侵袭数目显著少于联合用药组（P<0.05），anti-miR-433-3p+联合用药组细胞增殖抑制率和凋亡率显著低于联合用药组，细胞侵袭数目显著多于联合用药组（P<0.05）。miR-433-3p组SRC-WT荧光素酶活性显著低于miR-NC组（P<0.05）。si-SRC组细胞中SRC蛋白表达显著低于对照组（P<0.05），pcDNA-SRC组细胞中SRC蛋白表达显著高于对照组（P<0.05）；与pcDNA-SRC组相比，miR-433-3p+pcDNA-SRC组细胞中SRC蛋白表达显著降低（P<0.05）。si-SRC+联合用药组细胞增殖抑制率和凋亡率显著高于联合用药组，细胞侵袭数目显著少于联合用药组（P<0.05），pcDNA-SRC+联合用药组细胞增殖抑制率和凋亡率显著低于联合用药组，细胞侵袭数显著高于联合用药组（P<0.05）。与pcDNA-SRC+联合用药组相比，miR-433-3p+pcDNA-SRC+联合用药组细胞增殖抑制率和凋亡率显著降低（P<0.05），细胞侵袭数目显著升高（P<0.05）。结论 黄芩苷联合奥沙利铂可通过miR-433-3p靶向调控SRC抑制胃癌细胞的增殖和侵袭，诱导胃癌细胞凋亡。

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+pcDNASRC 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 pcDNASRC+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.

R285.5

上海市虹口区卫生健康委员会中医药科研课题（HKQ-ZYY-2020-21）