Department of Gynaecology, Xuzhou Maternal & Child Health Care Hospital, Xuzhou, Jiangsu, China.
J Cell Biochem. 2018 Apr;119(4):3058-3066. doi: 10.1002/jcb.26441. Epub 2018 Jan 2.
The present study was conducted to investigate the effects of RECK on cervical cancer cell migration and invasion to help understand relevant molecular mechanisms. QRT-PCR and western blot were respectively utilized to examine the transcriptional and translational levels of RECK in cervical cancer cell lines (HELA and C33A) and normal cell line (H8). After transfection with RECK overexpressing vectors, the expression of RECK mRNA, RECK and p53 signaling pathway-related proteins (p21, p53, bcl-2, and Bax) in cervical cancer cells were respectively examined using qRT-PCR and western blot. Cervical cancer cell migration after transfection was detected by wound healing assay and transwell assay. RECK expression was much lower in cervical cancer cell lines compared with normal cell line. Results of wound-healing assay results indicated that RECK could inhibit cervical cancer cell migration, and transwell assay results demonstrated that cell invasion was suppressed by RECK overexpression. Furthermore, western blot indicated that the overexpression of RECK could promote the activation of p53 signaling pathway by influencing related protein expression; whereas its inhibition by PFT-α could antagonize the effect of RECK on migrative and invasive abilities of cervical cancer cells. RECK could inhibit the migration and invasion of cervical cancer cells by activating p53 signaling pathway.
本研究旨在探讨 RE CK 对宫颈癌细胞迁移和侵袭的影响，以帮助理解相关的分子机制。我们分别采用 q RT-PCR 和 Western blot 检测了宫颈癌细胞系（H ELA 和 C33A）和正常细胞系（H8）中 RECK 的转录和翻译水平。转染 RECK 过表达载体后，采用 q RT-PCR 和 Western blot 分别检测宫颈癌细胞中 RECK m RNA、RECK 和 p53 信号通路相关蛋白（p21、p53、bcl-2 和 Bax）的表达。通过划痕愈合实验和 Transwell 实验检测转染后宫颈癌细胞的迁移。与正常细胞系相比，宫颈癌细胞系中 RECK 的表达水平较低。划痕愈合实验结果表明，RECK 可抑制宫颈癌细胞迁移，Transwell 实验结果表明，RECK 的过表达可抑制细胞侵袭。此外，Western blot 结果表明，通过影响相关蛋白的表达，RECK 的过表达可促进 p53 信号通路的激活；而 PFT-α 对其的抑制可拮抗 RECK 对宫颈癌细胞迁移和侵袭能力的影响。RECK 可通过激活 p53 信号通路抑制宫颈癌细胞的迁移和侵袭。
