目的 探討松果菊苷對皮膚鱗狀細(xì)胞癌A431細(xì)胞增殖及凋亡的影響及其作用機(jī)制。方法 體外培養(yǎng)人皮膚鱗狀細(xì)胞癌A431細(xì)胞，采用不同質(zhì)量濃度（12.5、25.0、50.0 μg/L）的松果菊苷處理A431細(xì)胞。將circ_0046264對照空載體（pcDNA）、circ_0046264過表達(dá)載體（pcDNA-circ_0046264）、miR-510寡核苷酸陰性對照（anti-miR-NC）、miR-510寡核苷酸抑制物（miR-510 inhibitor）分別轉(zhuǎn)染至A431細(xì)胞。將circ_0046264小分子干擾RNA（si-circ_0046264）、miR-510寡核苷酸模擬物（miR-510 mimic）分別轉(zhuǎn)染至A431細(xì)胞后加入50.0 μg/mL松果菊苷進(jìn)行處理。采用MTT法、平板克隆形成實(shí)驗(yàn)與流式細(xì)胞術(shù)分別檢測細(xì)胞增殖、克隆形成及凋亡率；qRT-PCR法與Western blotting法分別檢測circ_0046264、miR-510、阿片樣物質(zhì)結(jié)合蛋白（OPCML）的表達(dá)量；雙熒光素酶報告實(shí)驗(yàn)檢測circ_0046264與miR-510的靶向關(guān)系，以及miR-510與OPCML的靶向關(guān)系。結(jié)果 松果菊苷可濃度相關(guān)性地升高A431細(xì)胞增殖抑制率、凋亡率和circ_0046264、OPCML的表達(dá)量，并可降低miR-510的表達(dá)量，集落形成數(shù)減少（P<0.05）。轉(zhuǎn)染pcDNA-circ_0046264或miR-510 inhibitor后，OPCML蛋白表達(dá)顯著升高，細(xì)胞增殖抑制率和凋亡率顯著升高，集落形成數(shù)明顯減少（P<0.05）。circ_0046264可靶向調(diào)控miR-510的表達(dá)，OPCML是miR-510的靶基因；轉(zhuǎn)染si-circ_0046264或miR-510 mimic均可降低松果菊苷對A431細(xì)胞增殖及凋亡的作用。結(jié)論 松果菊苷可通過調(diào)節(jié)circ_0046264/miR-510/OPCML表達(dá)而抑制皮膚鱗狀細(xì)胞癌細(xì)胞增殖及誘導(dǎo)細(xì)胞凋亡。

Objective To investigate the effects of echinoside on proliferation and apoptosis of squamous cell carcinoma A431 cells and its mechanism. Methods Human squamous cell carcinoma A431 cells were cultured in vitro and treated with echinoside at different concentrations (12.5, 25.0, and 50.0 μg/). pc, pc-circ_0046264, anti-mi-, and mi-510 inhibitor were transfected into 431 cells, respectively. si-circ_0046264 and mi-510 mimic were transfected into 431 cells and 50.0 μg/m ecsinoside was added. assay, plate clonogenesis assay, and flow cytometry were used to detect cell proliferation, clonogenesis and apoptosis rates, respectively. he expression levels of circ_0046264, mi-510, and were detected by q-R and Western blotting method, respectively. The targeting relationship between circ_0046264 and miR-510, and the targeting relationship between miR-510 and OPCML were detected by dual-lucifase reporting assay. Results Ecinosin increased proliferation inhibition and apoptosis rate, circ_0046264 and OPCML expression, decreased miR-510 expression level, and decreased colony formation number (P < 0.05). After transfection with pcDNA-circ_0046264 or miR-510 inhibitor, OPCML protein expression, cell proliferation inhibition rate, and apoptosis rate were significantly increased, and colony formation number was significantly decreased (P< 0.05).circ_0046264 can regulate the expression of miR-510, and OPCML is the target gene of miR-510. Transfection with si-circ_0046264 or miR-510 mimic could reduce the proliferation and apoptosis of A431 cells. Conclusion Echinoside can inhibit proliferation and induce apoptosis of squamous cell carcinoma cells by regulating circ_0046264/miR-510/OPCML expression.

R285.5

河南省醫(yī)學(xué)科技攻關(guān)計(jì)劃項(xiàng)目（LHGJ20191162）