溶血磷脂酸对宫颈癌细胞生长增殖及MMP-2、Laminin5表达的影响

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目的:观察溶血磷脂酸对宫颈癌细胞株Siha生长增殖及对细胞表达MMP-2、Laminin5的影响,探讨溶血磷脂酸与宫颈癌发生发展的关系。方法:①以1μmol/L、5μmol/L、10μmol/L、15μmol/L的溶血磷脂酸和5 mg/L、10 mg/L、20mg/L、40 mg/L的多西环素体外干预人宫颈癌Siha细胞株。各组均于干预72 h后采用MTT法检测宫颈癌细胞活性。②以终浓度为10μmol/L的溶血磷脂酸和终浓度为40 mg/L的多西环素分别干预Siha细胞,各组培养72 h后提取总蛋白,采用Western blot检测Siha细胞MMP-2、Laminin5蛋白表达。结果:①MTT法检测溶血磷脂酸组的细胞抑制率随着药物浓度增加而逐渐下降,多西环素组的细胞抑制率随着药物浓度增加而逐渐增加(P<0.05);溶血磷脂酸和多西环素对Siha细胞的影响均呈浓度依赖关系。②溶血磷脂酸组MMP-2和Laminin5蛋白表达均显著高于空白对照组(P<0.01和P<0.05)。多西环素组MMP-2蛋白表达低于空白对照组,但无统计学意义(P>0.05);Laminin5蛋白含量显著降低(P<0.05)。结论:溶血磷脂酸对宫颈癌细胞株Siha生长增殖具有促进作用,并能增强细胞表达MMP-2、Laminin5;而多西环素明显抑制Siha细胞的生长增殖及MMP-2、Laminin5的表达。 OBJECTIVE: To investigate the effect of lysophosphatidic acid on the growth and proliferation of cervical cancer cell line Siha and on the expression of MMP-2 and Laminin5 in cervical cancer cells and explore the relationship between lysophosphatidic acid and the development of cervical cancer. Methods ①In vitro intervention with 1μmol / L, 5μmol / L, 10μmol / L, 15μmol / L lysophosphatidic acid and doxycycline 5 mg / L, 10 mg / L, 20 mg / L and 40 mg / L Cervical cancer Siha cell line. After 72 h of intervention, the activity of cervical cancer cells in each group was detected by MTT assay. (2) Siha cells were treated with lysophosphatidic acid at a final concentration of 10 μmol / L and doxycycline at a final concentration of 40 mg / L, and the total protein was extracted after 72 h of culture in each group. MMP-2, Laminin5 protein expression. Results: (1) The inhibition rate of MTT assay decreased gradually with the increase of drug concentration. The cell inhibition rate of doxycycline group increased gradually with the increase of drug concentration (P <0.05); the lysophosphatidic acid and Ciclopirox on Siha cells in a concentration-dependent manner. ② The protein expressions of MMP-2 and Laminin5 in lysophosphatidic acid group were significantly higher than those in blank control group (P <0.01 and P <0.05). The expression of MMP-2 protein in doxycycline group was lower than that in the blank control group, but there was no statistical significance (P> 0.05). The protein content of Laminin5 was significantly decreased (P <0.05). CONCLUSION: Lysophosphatidic acid can promote the growth of cervical cancer cell line Siha and enhance the expression of MMP-2 and Laminin5, while doxycycline significantly inhibits the growth of Siha cells and the expression of MMP-2 and Laminin5.
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