Sun Ren-Hu, Wang Guo-Bin, Li Jiang, Cui Jing
Department of Laparoscopic Center, Huazhong University of Science and Technology, Wuhan, Hubei, People's Republic of China.
Ai Zheng. 2009 Jul;28(7):708-13. doi: 10.5732/cjc.008.10786.
Chemokine receptor CCR7 is up-regulated in gastrointestinal carcinomas and is significantly associated with lymphatic invasion and lymph node metastasis. This study was to investigate the role and mechanism of CCL21/CCR7 in invasion of colorectal carcinoma cell line SW480.
The invasive capacity of SW480 cells was examined using Wound healing assay and Transwell assay. Expression of matrix metalloproteinase-9 (MMP-9) was measured by Western blot. SW480 cells were pre-incubated with CCL21 for 2 h before exposure to VP-16 (20 ng/mL). Cell proliferation was measured using MTT assay. Cell apoptosis was analyzed by flow cytometry and Hoechst33258 staining.
Compared to the control group, more cells in the CCL21 treatment group migrated into the gap at same time points; the count of SW480 cells penetrating through the membrane after the treatment of 100ng/mL CCL21 increased significantly [(113+/-7) vs. (48+/-4)] (P<0.05); and the relative expression of MMP-9 in the CCL21 treatment group was enhanced evidently [(0.83+/-0.02) vs. (0.38+/-0.01)] (P<0.05). Although CCL21 alone did not promote proliferation of SW480 cells, pre-incubation of cells with 100ng/mL CCL21 attenuated the inhibitory effect of VP-16 on proliferation of SW480 from 68.3% to 47.4%, and reduced the apoptotic rate from (65.2+/-5.2)% to (48.7+/-3.1)%.
CCL21 enhances the invasive ability of SW480 cells, induces MMP-9 expression, and promotes the survival of SW480 cells under the suboptimal circumstance in vitro.
趋化因子受体CCR7在胃肠道癌中表达上调，且与淋巴管浸润及淋巴结转移显著相关。本研究旨在探讨CCL21/CCR7在结肠癌细胞系SW480侵袭中的作用及机制。
采用伤口愈合试验和Transwell试验检测SW480细胞的侵袭能力。通过蛋白质印迹法检测基质金属蛋白酶-9（MMP-9）的表达。SW480细胞在暴露于VP-16（20 ng/mL）前用CCL21预孵育2小时。采用MTT试验检测细胞增殖。通过流式细胞术和Hoechst33258染色分析细胞凋亡。
与对照组相比，CCL21处理组在相同时间点有更多细胞迁移至间隙；100ng/mL CCL21处理后穿透膜的SW480细胞计数显著增加[（113±7）对（48±4）]（P<0.05）；CCL21处理组中MMP-9的相对表达明显增强[（0.83±0.02）对（0.38±0.01）]（P<0.05）。虽然单独的CCL21不促进SW480细胞增殖，但用100ng/mL CCL21预孵育细胞可将VP-16对SW480细胞增殖的抑制作用从68.3%减弱至47.4%，并使凋亡率从（65.2±5.2）%降低至（48.7±3.1）%。
CCL21增强SW480细胞的侵袭能力，诱导MMP-9表达，并在体外次优条件下促进SW480细胞存活。
