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目的:研究新疆阿魏乙酸乙酯提取物对人结肠腺癌Caco-2细胞迁移、侵袭的影响,并探讨其相关机制。方法:以Caco-2细胞为研究对象,用6.25,12.5 mg·L-1新疆阿魏乙酸乙酯提取物和顺铂分别作用于处于对数生长期的人结肠腺癌Caco-2细胞,并设空白组,用划痕愈合法观察0~48 h各组Caco-2细胞迁移情况,用transwell小室法计数72 h各组Caco-2细胞穿过基质膜个数,并计算出每个时间段Caco-2细胞迁移率和侵袭抑制率。采用荧光实时定量PCR法和蛋白免疫印迹法(Western blot)检测各组Caco-2细胞中磷酸酶(phosphatase and tensin homolog deleted on chromosome ten,PTEN),丝氨酸/苏氨酸蛋白激酶(serine/threonine,Akt),哺乳动物雷帕霉素靶蛋白(mammalina target of rapamgcin,m TOR)基因和蛋白的表达水平。结果:与空白组比较,新疆阿魏乙酸乙酯提取物和顺铂组在24,48 h细胞愈合程度变慢(P<0.05);72 h时穿过基质膜的细胞个数显著减少(P<0.01),各用药组PTEN基因表达水平明显升高,Akt,m TOR基因表达水平明显较低(P<0.05)。结论:新疆阿魏乙酸乙酯提取物可以显著抑制人结肠腺癌Caco-2细胞的迁移和侵袭,可能与提高PTEN基因表达水平有关。
OBJECTIVE: To study the effect of extract of Eruca sagittate on the migration and invasion of human colon adenocarcinoma Caco-2 cells and to explore the related mechanisms. Methods: The human colon adenocarcinoma Caco-2 cells were treated with 6.25, 12.5 mg · L-1 Xinjiang Efexel and Cisplatin respectively in Caco-2 cells. The blank group was established and the migration of Caco-2 cells in each group was observed by scratch healing method. The number of Caco-2 cells passing through the stroma was counted by transwell chamber method at each time point Caco-2 cell migration and invasion inhibition rate. The expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN), serine / threonine protein kinase (CIN) in each group were detected by real-time fluorescence quantitative PCR and Western blot. Akt), mammalian target protein of mammalian target of rapamycin (m TOR). Results: Compared with the blank group, the cells in the extract of E.globactex and the cisplatin group showed a slower healing at 24 and 48 h (P <0.05), and the number of cells crossing the stromal membrane at 72 h decreased significantly (P <0.01). The expression of PTEN gene in each treatment group was significantly increased, while the expression of Akt, m TOR gene was significantly lower (P <0.05). CONCLUSION: Eustoin extract can significantly inhibit the migration and invasion of human colon adenocarcinoma Caco-2 cells, which may be related to the increase of PTEN gene expression.