Januchowski Radosſaw, Wojtowicz Karolina, Sterzyſska Karolina, Sosiſska Patrycja, Andrzejewska Maſgorzata, Zawierucha Piotr, Nowicki Michaſ, Zabel Maciej
Department of Histology and Embryology, Poznaſ University of Medical Sciences, Poland.
Department of Histology and Embryology, Poznaſ University of Medical Sciences, Poland.
Int J Biochem Cell Biol. 2016 Sep;78:248-259. doi: 10.1016/j.biocel.2016.07.017. Epub 2016 Jul 19.
The high mortality of ovarian cancer patients results from the failure of treatment caused by the inherent or acquired chemotherapy drug resistance. It was reported that overexpression of aldehyde dehydrogenase A1 (ALDH1A1) in cancer cells can be responsible for the development of drug resistance. To add the high expression of the drug transporter proteins the ALDHA1 is considered as a molecular target in cancer therapy. Therefore, we analysed drug-resistant ovarian cancer cell lines according to ALDHA1 expression and the association with drug resistance. The expression of ALDH1A1, P-glycoprotein (P-gp) or breast cancer resistance protein (BCRP) was determined using a microarray and confirmed by Q-PCR, western blot and fluorescence analysis. ALDH1A1 activity was determined using an Aldefluor assay. The impact of all-trans retinoic acid (ATRA) and diethylaminobenzaldehyde (DEAB) on chemotherapy resistance was assessed by the MTT chemosensitivity assay. The most abundant expression of ALDH1A1 was noted in paclitaxel- and topotecan-resistant cell lines where two populations of ALDH-positive and ALDH-negative cells could be observed. Those cell lines also revealed the overexpression of P-gp and BCRP respectively, and were able to form spheres in non-adherent conditions. Pre-treatment with ATRA and DEAB reduced chemotherapy resistance in both cell lines. ATRA treatment led to downregulation of the ALDH1A1, P-gp and BCRP proteins. DEAB treatment led to downregulation of the P-gp protein and BCRP transcript and protein. Our results indicate that ALDH1A1-positive cancer cells can be responsible for drug resistance development in ovarian cancer. Developing more specific ALDH1A1 inhibitors can increase chemotherapy effectiveness in ovarian cancer.
卵巢癌患者的高死亡率源于内在或获得性化疗耐药导致的治疗失败。据报道，癌细胞中醛脱氢酶A1（ALDH1A1）的过表达可能导致耐药性的产生。加上药物转运蛋白的高表达，ALDHA1被认为是癌症治疗的一个分子靶点。因此，我们根据ALDHA1的表达及其与耐药性的关联分析了耐药卵巢癌细胞系。使用微阵列测定ALDH1A1、P-糖蛋白（P-gp）或乳腺癌耐药蛋白（BCRP）的表达，并通过Q-PCR、western印迹和荧光分析进行确认。使用Aldefluor测定法测定ALDH1A1活性。通过MTT化学敏感性测定评估全反式维甲酸（ATRA）和二乙氨基苯甲醛（DEAB）对化疗耐药性的影响。在对紫杉醇和拓扑替康耐药的细胞系中观察到ALDH1A1表达最为丰富，其中可以观察到ALDH阳性和ALDH阴性细胞群体。这些细胞系还分别显示出P-gp和BCRP的过表达，并且能够在非贴壁条件下形成球体。用ATRA和DEAB预处理可降低两种细胞系的化疗耐药性。ATRA处理导致ALDH1A1、P-gp和BCRP蛋白的下调。DEAB处理导致P-gp蛋白以及BCRP转录本和蛋白的下调。我们的结果表明，ALDH1A1阳性癌细胞可能是卵巢癌耐药性产生的原因。开发更具特异性的ALDH1A1抑制剂可以提高卵巢癌化疗的有效性。
