Shanghai Minimally Invasive Surgery Center, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, No. 197 Second Ruijin Road, Shanghai, China.
Dig Dis Sci. 2014 Sep;59(9):2153-62. doi: 10.1007/s10620-014-3138-y. Epub 2014 Apr 5.
To investigate the function of CC motif chemokine ligand 19 (CCL19) in colorectal cancer (CRC).
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot and immunohistochemistry were performed separately to detect the expression of CCL19 in colorectal carcinoma tissues. The expression of CCL19 and its receptor (CCR7) in CRC cell lines were screened by Western blot. SW620, SW1116 and LoVo cell lines were screened and processed with recombinant human CCL19 (rhCCL19) or si-CCL19 RNA. Cell proliferation assay and transwell assay were performed to evaluate the proliferation, migration and invasion of CRC cells, respectively. And the role of proangiogenesis was checked by endothelial tube formation assay.
qRT-PCR, Western blot and immunohistochemistry revealed that both CCL19 mRNA and protein were obviously expressed in a lower degree in CRC tissues than normal tissues (P < 0.01). The CCL19 expression correlated with tumor size (P = 0.03) and invasion depth (P = 0.04) in a negative manner and CCL19-positive patients had longer lifespans (P < 0.05). SW620 and SW1116 cells were screened as CCL19/CCR7 high-expression cells, while LoVo was selected as CCL19/CCR7 low-expression cell among seven CRC cell lines by Western blot. The proliferation, migration, invasion and proangiogenesis of SW620 and SW1116 cells were distinctly suppressed after they were stimulated by rhCCL19 (P < 0.05), and the data presented dose-dependency. Oppositely, these abilities were significantly enhanced after CCL19 gene was silenced (P < 0.05). However, the effects of rhCCL19 and si-CCL19 RNA on LoVo were not significant (P > 0.05).
Our research findings indicate that CCL19 may play a suppressive role in colorectal tumorigenesis.
研究 CC 基序趋化因子配体 19(CCL19)在结直肠癌(CRC)中的作用。
分别采用定量逆转录聚合酶链反应(qRT-PCR)、Western blot 和免疫组织化学方法检测结直肠癌组织中 CCL19 的表达。Western blot 筛选 CRC 细胞系中 CCL19 及其受体(CCR7)的表达。筛选 SW620、SW1116 和 LoVo 细胞系,并分别用重组人 CCL19(rhCCL19)或 si-CCL19 RNA 进行处理。细胞增殖试验和 Transwell 试验分别用于评估 CRC 细胞的增殖、迁移和侵袭能力,内皮管状形成试验用于检查促血管生成作用。
qRT-PCR、Western blot 和免疫组织化学结果显示,CRC 组织中 CCL19 mRNA 和蛋白的表达明显低于正常组织(P < 0.01)。CCL19 的表达与肿瘤大小(P = 0.03)和浸润深度(P = 0.04)呈负相关,CCL19 阳性患者的生存期更长(P < 0.05)。Western blot 筛选出 SW620 和 SW1116 细胞为 CCL19/CCR7 高表达细胞,而在 7 种 CRC 细胞系中 LoVo 细胞为 CCL19/CCR7 低表达细胞。rhCCL19 刺激 SW620 和 SW1116 细胞后,其增殖、迁移、侵袭和促血管生成能力明显受到抑制(P < 0.05),且呈剂量依赖性。相反,CCL19 基因沉默后,这些能力显著增强(P < 0.05)。然而,rhCCL19 和 si-CCL19 RNA 对 LoVo 的作用不明显(P > 0.05)。
我们的研究结果表明,CCL19 可能在结直肠肿瘤发生中发挥抑制作用。