[關(guān)鍵詞]
[摘要]
目的 探究黃芩苷聯(lián)合奧沙利鉑調(diào)控miR-433-3p/SRC對(duì)胃癌細(xì)胞增殖和侵襲的影響。方法 將SGC-7901細(xì)胞分為對(duì)照組、黃芩苷(100、200、300 μmol·L-1)組、奧沙利鉑(33 μmol·L-1)組,聯(lián)合用藥(300 μmol·L-1黃芩苷+33 μmol·L-1奧沙利鉑)組,miR-433-3p組、miR-NC組、anti-miR-433-3p組、anti-miR-NC組、pcDNA-SRC組、si-SRC組,miR-433-3p+聯(lián)合用藥組、anti-miR-433-3p+聯(lián)合用藥組、pcDNA-SRC+聯(lián)合用藥組、si-SRC+聯(lián)合用藥組、miR-433-3p+pcDNASRC+聯(lián)合用藥組。CCK-8檢測(cè)細(xì)胞增殖能力;Transwell小室法檢測(cè)細(xì)胞侵襲能力;流式細(xì)胞儀檢測(cè)細(xì)胞凋亡率;實(shí)時(shí)熒光定量PCR (qRT-PCR)檢測(cè)細(xì)胞中miR-433-3p表達(dá);雙熒光素酶報(bào)告檢測(cè)miR-433-3p和SRC的靶向關(guān)系;Westernblotting檢測(cè)細(xì)胞中SRC蛋白表達(dá)。結(jié)果 與對(duì)照組相比,黃芩苷(100、200、300 μmol·L-1)組、奧沙利鉑組、聯(lián)合用藥組細(xì)胞增殖抑制率、凋亡率和miR-433-3p表達(dá)顯著增加(P<0.05),細(xì)胞侵襲數(shù)目、SRC蛋白表達(dá)顯著減少(P<0.05);與黃芩苷300 μmol·L-1組、奧沙利鉑組相比,聯(lián)合用藥組細(xì)胞增殖抑制率、凋亡率顯著增加,細(xì)胞侵襲數(shù)目顯著減少;與黃芩苷300 μmol·L-1組比較,miR-433-3p表達(dá)顯著增加(P<0.05);與奧沙利鉑組比較,SRC蛋白表達(dá)顯著減少(P<0.05)。與對(duì)照組比較,miR-433-3p組細(xì)胞中miR-433-3p表達(dá)顯著升高,SRC蛋白表達(dá)顯著降低,anti-miR-433-3p組細(xì)胞中miR-433-3p表達(dá)顯著降低,SRC蛋白表達(dá)顯著升高(P<0.05);miR-433-3p+聯(lián)合用藥組細(xì)胞增殖抑制率和凋亡率顯著高于聯(lián)合用藥組,細(xì)胞侵襲數(shù)目顯著少于聯(lián)合用藥組(P<0.05),anti-miR-433-3p+聯(lián)合用藥組細(xì)胞增殖抑制率和凋亡率顯著低于聯(lián)合用藥組,細(xì)胞侵襲數(shù)目顯著多于聯(lián)合用藥組(P<0.05)。miR-433-3p組SRC-WT熒光素酶活性顯著低于miR-NC組(P<0.05)。si-SRC組細(xì)胞中SRC蛋白表達(dá)顯著低于對(duì)照組(P<0.05),pcDNA-SRC組細(xì)胞中SRC蛋白表達(dá)顯著高于對(duì)照組(P<0.05);與pcDNA-SRC組相比,miR-433-3p+pcDNA-SRC組細(xì)胞中SRC蛋白表達(dá)顯著降低(P<0.05)。si-SRC+聯(lián)合用藥組細(xì)胞增殖抑制率和凋亡率顯著高于聯(lián)合用藥組,細(xì)胞侵襲數(shù)目顯著少于聯(lián)合用藥組(P<0.05),pcDNA-SRC+聯(lián)合用藥組細(xì)胞增殖抑制率和凋亡率顯著低于聯(lián)合用藥組,細(xì)胞侵襲數(shù)顯著高于聯(lián)合用藥組(P<0.05)。與pcDNA-SRC+聯(lián)合用藥組相比,miR-433-3p+pcDNA-SRC+聯(lián)合用藥組細(xì)胞增殖抑制率和凋亡率顯著降低(P<0.05),細(xì)胞侵襲數(shù)目顯著升高(P<0.05)。結(jié)論 黃芩苷聯(lián)合奧沙利鉑可通過(guò)miR-433-3p靶向調(diào)控SRC抑制胃癌細(xì)胞的增殖和侵襲,誘導(dǎo)胃癌細(xì)胞凋亡。
[Key word]
[Abstract]
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.
[中圖分類號(hào)]
R285.5
[基金項(xiàng)目]
上海市虹口區(qū)衛(wèi)生健康委員會(huì)中醫(yī)藥科研課題(HKQ-ZYY-2020-21)