Zheng Jian-Bao, Qiao Li-Na, Sun Xue-Jun, Qi Jie, Ren Hai-Liang, Wei Guang-Bing, Zhou Pei-Hua, Yao Jian-Feng, Zhang Li, Jia Peng-Bo
Department of General Surgery, First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Second Department of Cardiovascular Medicine, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China.
Mol Med Rep. 2015 Sep;12(3):3409-3415. doi: 10.3892/mmr.2015.3838. Epub 2015 May 25.
Caudal‑related homeobox transcription factor 2 (CDX2) is a transcription factor, which is specifically expressed in the adult intestine. It is essential for the development and homeostasis of the intestinal epithelium and its functions as a tumor suppressor have been demonstrated in the adult colon. The present study aimed to examine the inhibitory effects of the overexpression of CDX2 on subcutaneously‑transplanted tumors, derived from LoVo colon cancer cells, in nude mice, and to provide experimental evidence for the biotherapy of colon cancer. A pEGFP‑C1‑CDX2 eukaryotic expression vector was transfected into the LoVo cells via lipofection, and LoVo cells stably‑expressing CDX2 (pEGFP‑C1‑CDX2 cells) were obtained using G418 selection. A nude mouse subcutaneously‑transplanted tumor model was established by inoculating the nude mice with the pEGFP‑C1‑CDX2 cells, and the effects of overexpression of CDX2 on transplanted tumor growth in the LoVo cells were observed. Western blotting results demonstrated that the protein expression of CDX2 in the LoVo cells was higher in the pEGFP‑C1‑CDX2 cell group, compared with that in the pEGFP‑C1 cell group and the untreated cell group. At 20 days post‑inoculation with either pEGFP‑C1‑CDX2 or pEGFP‑C1, the transplanted tumor masses were significantly lower in the pEGFP‑C1‑CDX2 group, compared with those in the pEGFP‑C1 and untreated groups. Immunohistochemistry revealed that the expression levels of CDX2 and matrix metalloproteinase‑2 (MMP‑2) were detected in each group, and the protein expression of CDX2 was increased in the tumor tissues from the nude mice in the pEGFP‑C1‑CDX2 group. However the expression of MMP‑2 was downregulated in the tumor tissues of the nude mice in the pEGFP‑C1‑CDX2 group. Taken together, these data suggested that pEGFP‑C1‑CDX2 cells exhibited suppressed tumor growth in vivo. Overexpression of CDX2 was observed in transplanted tumors in the pEGFP‑C1‑CDX2 group, and the gene expression of MMP‑2 was reduced. These results indicate that CDX2 inhibited the growth of colorectal tumor cells, possibly by downregulating the gene expression.
尾型相关同源框转录因子2(CDX2)是一种转录因子,在成年肠道中特异性表达。它对肠上皮的发育和稳态至关重要,并且其作为肿瘤抑制因子的功能已在成年结肠中得到证实。本研究旨在检测CDX2过表达对裸鼠体内源自LoVo结肠癌细胞的皮下移植瘤的抑制作用,并为结肠癌的生物治疗提供实验依据。通过脂质转染将pEGFP-C1-CDX2真核表达载体转染至LoVo细胞中,并使用G418筛选获得稳定表达CDX2的LoVo细胞(pEGFP-C1-CDX2细胞)。通过给裸鼠接种pEGFP-C1-CDX2细胞建立裸鼠皮下移植瘤模型,并观察CDX2过表达对LoVo细胞中移植瘤生长的影响。蛋白质印迹结果表明,与pEGFP-C1细胞组和未处理细胞组相比,pEGFP-C1-CDX2细胞组中LoVo细胞中CDX2的蛋白表达更高。在接种pEGFP-C1-CDX2或pEGFP-C1后20天,与pEGFP-C1组和未处理组相比,pEGFP-C1-CDX2组中的移植瘤块明显更小。免疫组织化学显示,在每组中检测到CDX2和基质金属蛋白酶-2(MMP-2)的表达水平,并且pEGFP-C1-CDX2组裸鼠肿瘤组织中CDX2的蛋白表达增加。然而,pEGFP-C1-CDX2组裸鼠肿瘤组织中MMP-2的表达下调。综上所述,这些数据表明pEGFP-C1-CDX2细胞在体内表现出抑制肿瘤生长的作用。在pEGFP-C1-CDX2组的移植瘤中观察到CDX2过表达,并且MMP-2的基因表达降低。这些结果表明,CDX2可能通过下调基因表达来抑制结直肠肿瘤细胞的生长。