目的 探究光甘草定（GLA）對(duì)食管癌細(xì)胞增殖、凋亡、侵襲、遷移的影響并分析其是否與調(diào)節(jié)Janus蛋白酪氨酸激酶2（JAK2）/信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子3（STAT3）通路有關(guān)。方法 MTT法檢測(cè)0、10、20、40、80、160、320 μmol·L^-1GLA對(duì)人食管癌細(xì)胞KYSE150存活率的影響，并計(jì)算半數(shù)抑制濃度（IC₅₀）。確定80、40、20 μmol·L^-1為后續(xù)實(shí)驗(yàn)GLA濃度，并設(shè)置對(duì)照組、GLA（80 μmol·L^-1）＋Colivelin（0.5 μmol·L^-1，JAK2/STAT3通路激活劑）組、GLA（80 μmol·L^-1）＋baricitini （10 μmol·L^-1，JAK2抑制劑）組，CCK8法檢測(cè)細(xì)胞活力，集落形成實(shí)驗(yàn)檢測(cè)細(xì)胞克隆能力，流式細(xì)胞儀檢測(cè)細(xì)胞凋亡，Transwell實(shí)驗(yàn)檢測(cè)細(xì)胞侵襲，傷口愈合實(shí)驗(yàn)檢測(cè)細(xì)胞遷移，Westren blotting法檢測(cè)JAK2/STAT3通路及增殖、遷移蛋白表達(dá)。建立食管癌裸鼠模型，隨機(jī)分為模型組和GLA（50 mg·kg^-1）組，每天ig給藥1次，治療3周后處死小鼠，分析腫瘤質(zhì)量、體積，免疫組化染色分析組織中JAK2、STAT3陽(yáng)性表達(dá)。結(jié)果 與對(duì)照組比較，GLA組凋亡率顯著升高（P＜0.05），細(xì)胞活力、細(xì)胞克隆數(shù)、侵襲數(shù)、遷移率顯著降低（P＜0.05）。與GLA 80 μmol·L^-1組比較，GLA＋Colivelin組凋亡率顯著降低（P＜0.05），細(xì)胞活力、細(xì)胞克隆數(shù)、侵襲數(shù)、遷移率顯著升高（P＜0.05）；GLA＋baricitini組細(xì)胞凋亡率顯著升高（P＜0.05），活力、細(xì)胞克隆數(shù)、侵襲數(shù)、遷移率顯著降低（P＜0.05）。與對(duì)照組比較，GLA組細(xì)胞Bax蛋白表達(dá)顯著升高（P＜0.05），p-JAK2、p-STAT3、c-MYC、Ki67、基質(zhì)金屬蛋白酶9 （MMP-9）表達(dá)顯著降低（P＜0.05）。與GLA 80 μmol·L^-1組比較，GLA＋Colivelin組Bax蛋白表達(dá)顯著降低（P＜0.05），p-JAK2、p-STAT3、c-MYC、Ki67、MMP-9蛋白表達(dá)顯著升高（P＜0.05）；GLA＋baricitini組Bax蛋白表達(dá)顯著升高（P＜0.05），p-JAK2、p-STAT3、c-MYC、Ki67、MMP-9蛋白表達(dá)顯著降低（P＜0.05）。與模型組裸鼠比較，GLA組腫瘤質(zhì)量和體積顯著降低（P＜0.05），JAK2、STAT3陽(yáng)性表達(dá)率顯著降低（P＜0.05）。結(jié)論 GLA可能抑制JAK2/STAT3通路，進(jìn)而抑制食管癌細(xì)胞增殖、侵襲、遷移，并促進(jìn)凋亡。

Objective To investigate the effects of glabridin (GLA) on the proliferation, apoptosis, invasion, and migration of esophageal cancer cells and analyze whether it is related to the regulation of the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway. Methods MTT assay was used to detect the cell survival rates of GLA intervention at 0, 10, 20, 40, 80, 160, and 320 μmol·L^-1, and IC₅₀ was calculated. 80, 40, and 20 μmol·L^-1 were determined as the doses for subsequent experiments in the GLA group, and control group, GLA (80 μmol·L^-1)+Colivelin (0.5 μmol·L^-1, JAK2/STAT3 pathway activator) group, GLA (80 μmol·L^-1) +baricitini (10 μmol·L^-1, JAK2 inhibitor) group were set up. CCK8 method was applied to detect cell viability, colony formation experiment was applied to detect cell cloning ability, transwell experiment was applied to detect cell invasion, the wound healing experiment was applied to detect cell migration, and immunoblotting was applied to detect the JAK2/STAT3 pathway and the expression of proliferation and migration proteins. A nude mouse model of esophageal cancer was established and randomly separated into a model group and a GLA (50 mg·kg^-1) group. Administer ig once a day, euthanize mice after three weeks of treatment, analyze tumor mass and volume, and perform immunohistochemical staining to analyze JAK2 and STAT3 positive expression in tissues. Results Compared with the control group, the apoptosis rate of the GLA group increased, while the cell viability, cell clone number, invasion number, and migration rate decreased (P < 0.05). Compared with the GLA 80 μmol·L^-1 group, the cell viability, number of cell clones, number of invasions, and migration rate in GLA+Colivelin group increased, while the apoptosis rate decreased (P < 0.05), the apoptosis rate of cells in the GLA+baricitinib group increased, while the vitality, number of cell clones, number of invasions, and migration rate decreased (P < 0.05). Compared with control group, the expression of Bax in cells of the GLA group increased, while the expression of p-JAK2, p-STAT3, c-MYC, Ki67, and MMP-9 decreased (P < 0.05). Compared with the GLA 80 μmol·L^-1 group, the expression of p-JAK2, p-STAT3, c-MYC, Ki67, and MMP-9 in the GLA+Colivelin group increased, while the expression of Bax decreased (P < 0.05); the expression of Bax in the GLA+ baricitinib group increased, while the expression of p-JAK2, p-STAT3, c-MYC, Ki67, and MMP-9 decreased (P < 0.05). Compared with the nude mice in model group, the tumor mass and volume in the GLA group decreased (P < 0.05), and the positive expression rates of JAK2 and STAT3 in the GLA group decreased (P < 0.05). Conclusion GLA can inhibit the JAK2/STAT3 pathway, thereby inhibiting the proliferation, invasion, migration, and promoting apoptosis of esophageal cancer cells.

R285.5

河南省教育科學(xué)“十四五”規(guī)劃項(xiàng)目（2021YB0515）