1 Department of Gynecology, Fujian Provincial Hospital, Fujian Provincial Clinical Medical College, Fujian Medical University , Fuzhou, Fujian, China .
2 Department of Pathology, Fujian Provincial Hospital, Fujian Provincial Clinical Medical College, Fujian Medical University , Fuzhou, Fujian, China .
Cancer Biother Radiopharm. 2017 May;32(4):139-146. doi: 10.1089/cbr.2016.2153.
Abnormal autophagy regulation affects the chemoresistance of ovarian cancer, during which the circadian gene clock may play a major role. In this study, RNA interference plasmid pSUPER-Clock and overexpression plasmid pcDNA3.1-Clock of CLOCK were used to stably transfect the SKOV3/DDP cells by lipofection. Upon screening, the in vitro transfected cell lines with pSUPER-Clock, the autophagy level, and G/G phase cells were significantly reduced, and the expression levels of Clock, LC3, P-gp, and MRP2 were inhibited. In contrast, the autophagy level and G/G phase cells in cell lines transfected with pcDNA3.1-Clock were significantly increased, and the expressions of Clock, LC3, P-gp, and MRP2 were enhanced. In comparison with the untransfected control group showed the percentage of apoptotic cells in SKOV3/DDP cell lines of Clock interfering expression group after cisplatin treatment was significantly increased while the survival was substantially reduced. These results indicated that inhibiting the circadian gene Clock expression can reverse the cisplatin resistance of ovarian cancer SKOV3/DDP cell lines by affecting the protein expression of drug resistance genes during which autophagy plays an important role. The CLOCK gene may be designated as a novel candidate for targeted gene therapy in drug-resistant ovarian cancer.
异常的自噬调控会影响卵巢癌的化疗耐药性，而昼夜节律基因时钟可能在此过程中发挥主要作用。在这项研究中，我们使用 RNA 干扰质粒 pSUPER-Clock 和过表达质粒 pcDNA3.1-Clock 对 SKOV3/DDP 细胞进行脂质体转染以实现稳定转染。经过筛选，pSUPER-Clock 体外转染细胞系中的自噬水平和 G/G 期细胞明显减少，Clock、LC3、P-gp 和 MRP2 的表达水平受到抑制。相反，pcDNA3.1-Clock 转染细胞系中的自噬水平和 G/G 期细胞明显增加，Clock、LC3、P-gp 和 MRP2 的表达水平增强。与未经转染的对照组相比，Clock 干扰表达组 SKOV3/DDP 细胞系在顺铂处理后的凋亡细胞比例明显增加，而存活率则显著降低。这些结果表明，抑制昼夜节律基因 Clock 的表达可以通过影响耐药基因的蛋白表达来逆转卵巢癌 SKOV3/DDP 细胞系对顺铂的耐药性，而自噬在此过程中发挥重要作用。CLOCK 基因可能被指定为耐药性卵巢癌靶向基因治疗的新候选基因。
