目的 探讨南蛇藤提取物（COE）联合miR-302对人食管癌细胞增殖、侵袭和迁移的影响及对PI3K/Akt信号通路的调控作用。方法 实时荧光定量PCR（qRT-PCR）检测人正常食管上皮细胞Het-1A及不同食管癌细胞株中miR-302表达情况。将miR-302 mimic和阴性对照mimic control转染至人食管癌TE-1细胞中，qRT-PCR检测质粒转染后TE-1细胞中miR-302的表达情况。单独或联合使用COE作用TE-1细胞，CCK-8实验检测TE-1细胞增殖情况，Transwell实验检测TE-1细胞侵袭和迁移能力，Western blotting分析PI3K/Akt信号通路中相关蛋白表达情况。结果 食管癌细胞株中miR-302的表达明显低于正常食管上皮细胞（P<0.05）。转染miR-302 mimic能够有效提高TE-1细胞中miR-302的表达（P<0.05）。单独使用COE或过表达miR-302可抑制食管癌TE-1细胞增殖、侵袭和迁移（P<0.05），下调PI3K和p-Akt蛋白表达；二者联合使用对TE-1细胞增殖、侵袭和迁移抑制及对PI3K和p-Akt蛋白表达下调作用更显著（P<0.05）。结论 COE联合miR-302可协同抑制食管癌TE-1细胞增殖、侵袭和迁移，其作用机制可能与抑制PI3K/Akt信号通路的激活有关。

Objective To investigate the effects of the combination of Celastrus orbiculatus extracts and miR-302 on proliferation, invasion and migration of human esophageal cancer cells and the regulation of PI3K/Akt signaling pathway. Methods Quantitative RT-PCR was used to detect the expression of miR-302 in human normal esophageal epithelial cells Het-1A and different esophageal cancer cell lines. The miR-302 mimic and negative control mimic control were transfected into human esophageal cancer TE-1 cells, and qPCR was used to detect the expression of miR-302 in TE-1 cells after plasmid transfection. TE-1 cells were treated with C. orbiculatus extracts alone and combination treatment. The proliferation of TE-1 cells was detected by CCK-8 assay. The invasion and migration of TE-1 cells were detected by Transwell assay. Western blot analysis of the expression of related protein in the PI3K/Akt signaling pathway was carried out. Results The expression of miR-302 in esophageal cancer cell lines was significantly lower than that in esophageal epithelial cells (P < 0.05). Transfection of miR-302 mimic could effectively increase the expression of miR-302 in TE-1 cells (P < 0.05). The use of C. orbiculatus extracts alone or overexpression of miR-302 inhibited proliferation, invasion and migration of esophageal cancer TE-1 cells (P < 0.05), down-regulated PI3K and p-Akt protein expression; Combination treatment had more significant effect on inhibiting proliferation, invasion and migration of TE-1 cells and down-regulating protein expression of PI3K and p-Akt (P < 0.05). Conclusion Celastrus orbiculatus extracts combined with miR-302 can synergistically inhibit the proliferation, invasion, and migration of esophageal cancer TE-1 cells, and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway activation.

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