Xue Jun, Wu Xueliang, Qu Ming, Guo Fei, Han Lei, Sun Guangyuan, Yuan Zelong, Fan Shuang, Li Tian
Department of General Surgery, First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China.
School of Basic Medicine, The Fourth Military Medical University, 169 Changle West Road, Xi'an 710032, China.
Evid Based Complement Alternat Med. 2020 Feb 27;2020:5978131. doi: 10.1155/2020/5978131. eCollection 2020.
To investigate the effect of Runt-associated transcription factor 3 (RUNX3) on the invasion and metastasis of human colon cancer HT-29 cells and to preliminarily explore the mechanism of its anticancer effect.
The RUNX3 plasmid vector was transfected into human colon cancer HT-29 cells by liposome-mediated transfection, while the empty vector and the blank group were used as the control group. After Geneticin (G418) screening, HT-29 cells with stable expression of RUNX3 gene were obtained. The expressions of mRNA and proteins of RUNX3 and metalloproteinases (MMP)-2/9 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. Cell proliferation was determined by MTT assay. The effect of RUNX3 on invasion and metastasis of HT-29 cells was evaluated by scratch injury assay, Transwell chamber, and Matrigel invasion model.
RUNX3 was expressed stably in HT-29 cells after transfection. The expressions of RUNX3 mRNA and proteins in the experimental group were significantly higher than those in the blank/empty vector groups. Meanwhile, the expressions of MMP-2/9 mRNA and proteins in the observation group were significantly lower than those in the blank group and the empty vector group. The proliferation and migration ability in the experimental group was significantly lower than blank/empty vector groups from the third day. Transwell chamber experiment and Matrigel invasion assay showed that the number of Transwell cells was decreased significantly than blank/empty vector groups, but no difference was found between the blank group and the empty vector group.
RUNX3 can inhibit the invasion and metastasis of human colon cancer HT-29 cells, and the mechanism may be related to decreased expression of MMP-2 and MMP-9.
探讨 runt 相关转录因子 3（RUNX3）对人结肠癌 HT - 29 细胞侵袭和转移的影响，并初步探讨其抗癌作用机制。
采用脂质体介导的转染方法将 RUNX3 质粒载体转染至人结肠癌 HT - 29 细胞，同时将空载体和空白组作为对照组。经 Geneticin（G418）筛选，获得 RUNX3 基因稳定表达的 HT - 29 细胞。采用逆转录 - 聚合酶链反应（RT - PCR）和蛋白质免疫印迹法检测 RUNX3 及基质金属蛋白酶（MMP）-2/9 的 mRNA 和蛋白表达。采用 MTT 法测定细胞增殖情况。通过划痕实验、Transwell 小室和基质胶侵袭模型评估 RUNX3 对 HT - 29 细胞侵袭和转移的影响。
转染后 RUNX3 在 HT - 29 细胞中稳定表达。实验组 RUNX3 的 mRNA 和蛋白表达明显高于空白/空载体组。同时，观察组中 MMP - 2/9 的 mRNA 和蛋白表达明显低于空白组和空载体组。从第三天起，实验组的增殖和迁移能力明显低于空白/空载体组。Transwell 小室实验和基质胶侵袭实验显示，Transwell 细胞数量明显少于空白/空载体组，但空白组和空载体组之间无差异。
RUNX3 可抑制人结肠癌 HT - 29 细胞的侵袭和转移，其机制可能与 MMP - 2 和 MMP - 9 的表达降低有关。
