缺氧对人乳腺癌细胞系MCF-7生物学行为的影响

目的:研究缺氧对人乳腺癌细胞系MCF-7的生物学行为,增殖、侵袭能力的影响。通过影响缺氧诱导因子1α(hypoxic-induciblefactor1α,HIF-1α)的表达,检验其在这一过程中的作用,并进一步探讨作用机制。方法:慢病毒感染细胞,干扰乳腺癌细胞系MCF-7中HIF-1α的表达,得到HIF-1α表达正常的乳腺癌细胞系MCF-7-NC和HIF-1α表达受干扰的细胞系MCF-7-HIF△。分别低氧培养及化学药物诱导缺氧,用MTT法检测2组细胞系在缺氧和正常培养条件下的增殖能力,ANOVA检验检测其增殖能力的区别。Transwell实验检验缺氧和正常培养条件下人乳腺癌细胞的侵袭能力,计数通过Transwell小室的细胞,ANOVA检验检测细胞侵袭能力的区别。共聚焦免疫荧光法检测缺氧后2组乳腺癌细胞的上皮和间质标记物的表达,检验细胞系是否发生了上皮-间质转化(epithelial-mesenchymaltransition,EMT)。结果:MTT实验结果:正常表达HIF-1α的MCF-7-NC的增殖能力在缺氧后与正常培养条件下相比明显减弱,其差别具有统计学意义(P<0.05)。HIF-1α表达受干扰的MCF-7-HIF△的增殖能力在缺氧和正常培养条件下相比无显著区别。Transwell实验检测细胞的侵袭能力,缺氧后正常表达HIF-1α的MCF-7-NC的侵袭能力较正常培养的细胞明显增强,其差别具有统计学意义(P<0.05)。HIF-1α表达受干扰的MCF-7-HIF△侵袭能力在缺氧和正常培养下无显著区别。缺氧后,正常表达HIF-1α的细胞系MCF-7-NC发生上皮-间质转化(epithelial-mesenchymaltransition,EMT)。共聚焦荧光染色显示:在诱导缺氧后,正常表达HIF-1α的MCF-7-NC细胞极性发生变化,呈长梭形改变,上皮标志物人广谱角蛋白(pan-cytokeratin,P-CK)表达下调,间质标志物人α平滑肌动蛋白(α-SMA)表达上调;而HIF-1α表达受干扰的MCF-7-HIF△无明显EMT变化。结论:在体外条件下,缺氧可以引起乳腺癌细胞系MCF-7增殖能力减弱、侵袭能力增强,可以诱导乳腺癌细胞MCF-7发生EMT,干扰缺氧诱导因子亚基HIF-1α的表达,则缺氧对乳腺癌细胞增殖、侵袭的影响消失。

The anti - proliferation and anti - invasion effect of astragaloside IV on liver cancer cell line in vitro and its mechanism

Objective：To investigate the anti - Proliferation and anti - invasion effect of astragaloside IV on liver cancer cell line MeCC97 - e in vitro. Methods：MeCC97 - e cells were exPosed to different concentrations of astra-galoside IV（A - IV）for 24h,48h and 72h resPectively. The Proliferation was detected by 4 - methyl - teerazolium （MTT）. Transwell assay was aPlied to detect the inhibitory effect of A - IV on MeCC97 - e cell invasion. MeCC97- e cells were treated by 100μg/ ml A - IV for 48 hours. Then the exPressive changes for matrix metalloProteinase -2,9（MMP - 2,9）mRNA and Protein in MeCC97 - e cells were detected by real - time PCR and Western bolt re-sPectively. The contents of MMP - 2,9 in culture suPematant of MeCC97 - e cells were measured by gelatin zymog-raPhy. Results：MTT showed that A - IV had no significantly inhibitoty effect on MeCC97 - e cells with any concen-trations and times. The result of Transwell assay showed that A - IV could inhibit the invasion of MeCC97 - e cells in vitro. The exPressions of MMP - 2,9 mRNA and Protein reduced after treatment by 100μg/ ml A - IV for 48 hours. And the contents of MMP - 2,9 in culture suPematant of MeCC97 - e cells reduced. Conclusion：A - IV has no ob-vious inhibitory effect on the Proliferation of MeCC97 - e cells,but a strong inhibitory action on the invasion of Me-CC97 - e cells in vitro.