目的 探討香葉醇調(diào)控烷基化修復(fù)蛋白B同源物5（ALKBH5）對乳腺癌細(xì)胞上皮間質(zhì)轉(zhuǎn)化的影響。方法 以0、6.25、12.5、25、50、100、200 μg/mL香葉醇分別處理人乳腺癌MCF-7細(xì)胞24 h，篩選合適的香葉醇處理劑量。MCF-7細(xì)胞分為對照組、香葉醇（25、50、100 μg/mL）組、香葉醇＋空白質(zhì)粒組、香葉醇＋ALKBH5組，5-乙炔基-2'脫氧尿嘧啶核苷（EdU）染色、CCK-8法檢測細(xì)胞增殖；Western blotting檢測細(xì)胞中ALKBH5、神經(jīng)鈣黏蛋白（N-cadherin）、波形蛋白（Vimentin）、E-鈣黏蛋白（E-cadherin）蛋白；Transwell、劃痕實(shí)驗(yàn)分別檢測細(xì)胞侵襲、遷移；qRT-PCR檢測細(xì)胞中基質(zhì)金屬蛋白酶（MMP）-9、遷移侵襲增強(qiáng)子（MIEN1）、MMP-2 mRNA表達(dá)。結(jié)果 與0 μg/mL香葉醇比較，6.25、12.5、25、50、100、200 μg/mL香葉醇作用后的MCF-7細(xì)胞活力均顯著降低（P＜0.05），選取香葉醇質(zhì)量濃度為25、50、100 μg/mL作為后續(xù)處理濃度。與對照組比較，香葉醇25、50、100 μg/mL組MCF-7細(xì)胞EdU陽性率、A₄₅₀值、ALKBH5、N-cadherin、Vimentin蛋白表達(dá)、細(xì)胞侵襲數(shù)、劃痕愈合率及MMP-9、MIEN1、MMP-2 mRNA表達(dá)降低，E-cadherin蛋白表達(dá)升高（P＜0.05）。與香葉醇100 μg/mL組、香葉醇＋空白質(zhì)粒組比較，香葉醇＋ALKBH5組MCF-7細(xì)胞EdU陽性率、A₄₅₀值、ALKBH5、N-cadherin、Vimentin蛋白表達(dá)、細(xì)胞侵襲數(shù)、劃痕愈合率及MMP-9、MIEN1、MMP-2 mRNA表達(dá)升高，E-cadherin蛋白表達(dá)降低（P＜0.05）。結(jié)論 香葉醇可能通過下調(diào)ALKBH5抑制MCF-7細(xì)胞上皮間質(zhì)轉(zhuǎn)化，進(jìn)而減弱細(xì)胞侵襲與遷移能力。

Objective To investigate the effect of geraniol on epithelial mesenchymal transition of breast cancer cells by regulating ALKBH5. Methods Human breast cancer MCF-7 cells were treated with 0, 6.25, 12.5, 25, 50, 100, 200 μg/mL geraniol for 24 hours, and the appropriate dosage of geraniol was screened. MCF-7 cells were assigned into control group, geraniol (25, 50, 100 μg/mL) group, geraniol + blank plasmid set group, and geraniol + ALKBH5 group. EdU staining and CCK-8 method were used to detect cell proliferation. Western blotting was used to detect ALKBH5, N-cadherin, Vimentin, and E-cadherin proteins in cells. Transwell and scratch experiments were used to detect cell invasion and migration, respectively. QRT-PCR was used to measure the mRNA expression of MMP-9, MIEN1, and MMP-2 in cells. Results Compared with 0 μg/mL geraniol, the activity of MCF-7 cells after treatment with 6.25, 12.5, 25, 50, 100, 200 μg/mL geraniol was significantly decreased (P < 0.05). The concentration of geraniol was 25, 50, 100 μg/mL as the subsequent treatment concentration. Compared with control group, the EdU positive rate, A₄₅₀ value, ALKBH5, N-cadherin, Vimentin protein, cell invasion number, scratch healing rate, and MMP-9, MIEN1, MMP-2 mRNA expression of MCF-7 cells in the geraniol 25, 50, 100 μg/mL groups were reduced, while E-cadherin protein were increased (P < 0.05). Compared with the geraniol 100 μg/mL group and geraniol + blank plasmid set group, the EdU positive rate, A₄₅₀ value, ALKBH5, N-cadherin, Vimentin protein, cell invasion number, scratch healing rate, and MMP-9, MIEN1, MMP-2 mRNA expression of MCF-7 cells in geraniol + ALKBH5 group were increased, while E-cadherin protein were decreased (P < 0.05). Conclusion Geraniol may inhibit epithelial mesenchymal transition in MCF-7 cells by downregulating ALKBH5, thereby weakening cell invasion and migration abilities.

R979.1

河南省醫(yī)學(xué)科技攻關(guān)計劃聯(lián)合共建項目（LHGJ20220783，LHGJ20220784）