Papachristopoulou Georgia, Talieri Maroulio, Scorilas Andreas
Department of Biochemistry and Molecular Biology, University of Athens, 15701, Panepistimiopolis, Athens, Greece.
Tumour Biol. 2013 Feb;34(1):369-78. doi: 10.1007/s13277-012-0558-1. Epub 2012 Oct 20.
Given that 1.3 million new cases of breast cancer are universally registered among women and approximately 36 % of the patients die annually, the revelation of new predictive markers for treatment efficiency is of vital importance. Recently, our group has depicted that KLK4, KLK5, and KLK14 are differentially expressed in breast carcinoma. The objective of this study was to determine and investigate the expression pattern of the KLK4, KLK5, and KLK14 genes in breast cancer cells after treatment with established chemotherapeutic agents. We evaluated these genes' expression after treatment of the BT-20 cells with epirubicin, docetaxel and methotrexate, determining their cytotoxic effect by MTT and trypan blue assays. The relative quantification of genes' mRNA levels was performed by using the SYBR Green® chemistry, and the HPRT1 served as an endogenous control gene. The drugs triggered apoptosis in treated cells and induced deregulations in the expression of the above KLKs. The most significant alterations were a 12-fold and tenfold increase of KLK5 in docetaxel and methotrexate-treated cells, respectively, while the KLK4 levels decreased by ten-fold in epirubicin, five-fold in docetaxel and twenty-fold in methotrexate treated-cells, compared to the untreated ones. In the case of KLK14 levels, a twofold increase in epirubicin and threefold decrease in methotrexate-treated cells were observed. Present significant alterations in the expression pattern of KLK4, KLK5, and KLK14 could comprise an initial stage for predicting chemotherapy response in breast cancer and should be further investigated as predictive markers in the future.
鉴于全球每年有130万例新发乳腺癌病例,且约36%的患者每年死亡,揭示新的治疗效果预测标志物至关重要。最近,我们的研究小组发现KLK4、KLK5和KLK14在乳腺癌中存在差异表达。本研究的目的是确定并研究经既定化疗药物治疗后乳腺癌细胞中KLK4、KLK5和KLK14基因的表达模式。我们用表柔比星、多西他赛和甲氨蝶呤处理BT - 20细胞后评估这些基因的表达,并通过MTT和台盼蓝试验确定其细胞毒性作用。使用SYBR Green®化学方法对基因的mRNA水平进行相对定量,HPRT1作为内参基因。这些药物在处理后的细胞中引发凋亡,并诱导上述KLKs表达失调。最显著的变化是,多西他赛和甲氨蝶呤处理的细胞中KLK5分别增加了12倍和10倍,而与未处理细胞相比,表柔比星处理的细胞中KLK4水平降低了10倍,多西他赛处理的细胞中降低了5倍,甲氨蝶呤处理的细胞中降低了20倍。在KLK14水平方面,观察到表柔比星处理的细胞中增加了2倍,甲氨蝶呤处理的细胞中降低了3倍。KLK4、KLK5和KLK14表达模式目前的显著变化可能是预测乳腺癌化疗反应的初始阶段,未来应作为预测标志物进一步研究。