干扰DHRS9表达对三阴乳腺癌恶性生物学行为的调控作用研究

Regulation Effect of Interference DHRS9 on Malignant Biological Behaviour in Patients with Triple-negative Breast Cancer

目的探究脱氢酶/还原酶家族9(DHRS9)对三阴乳腺癌细胞干性相关基因的调控作用,并进一步分析其对癌细胞诸多恶性生物学行为的影响。方法应用Lipofectamine 2000转染试剂将sh DHRS9质粒及绿色荧光蛋白分别转染至三阴乳腺癌MDA-MB-231细胞中(分别设为实验组和对照组),应用划痕实验、Transwell实验检测细胞迁徙能力,运用免疫印迹法及实时荧光定量多聚酶链式反应(PCR),分别在蛋白及基因水平测定MDA-MB-231细胞中DHRS9表达量的变化;同时应用实时荧光定量PCR实验、划痕实验、Transwell实验及成球实验,观察下调DHRS9表达后对两组细胞干性相关基因及侵袭转移等恶性生物学行为的影响。结果实验组下调DHRS9表达后,CD133、ABCG2、CD44干性相关基因表达量升高(P<0.05),乳腺癌细胞侵袭迁移能力下降(P<0.05),复发转移能力及自我更新能力明显增强(P<0.05)。结论 DHRS9作为抑癌基因可能通过抑制三阴乳腺癌细胞干性相关基因表型,干预癌细胞恶性生物学行为,其可作为三阴乳腺癌的潜在抑癌分子标志物。

Objective To investigate regulation effect of dehydrogenase/reductase family 9 （DHRS9） on triple-negative breast cancer stem ceils related genes, and to further analyze its effects on malignant biological behaviour of cancer cells. Methods shDHRS9 plasmid （experimental group） and green fluorescent protein （GFP group） were respectively trausfected into cell line MDA-MB-231 cells in triple-negative breast cancer using Lipofectamine 2000 oligofectamine reagent. Migration ability of cells was detected using scuffing and Transwell experiments. Changes of DHRS9 expression were observed by Western blotting and real-time fluorescent quantitation polymerase chain reaction （PCR） respectively on proteic and genetic levels. Effects of down-regulated DHRS9 on MDA-MB-231 stem cells related gene and negative influence of malignant biological behaviour such as invasion and metastasis were observed by real-time fluorescent quantitatiou PCR, scuffing, Transwell and bailing experiments synchronously. Results In experimental group after down-regulated DHRS9 expression, expressions related CD133, CD90, ABCG2 and CD44 genes were increased （ P 〈 0.05 ） , abilities of invasion and metastasis of breast cancer cells were decreased （ P 〈 0.05 ） , and abilities of recurrence, metastasis and self-renewal were significantly improved （ P 〈 0.05 ）. Conclusion Anti-oncogene DHRS9 may interfere in malignant biological behaviour in triple-negative breast cancer cells by inhibiting gene phenotype of related stem ceils, and therefore it can be used as a potential tumor suppressor molecular marker.