Department of Life Sciences, Nanjing Normal University, Jiangsu Province, China.
Exp Biol Med (Maywood). 2013 Aug 1;238(8):903-12. doi: 10.1177/1535370213493706. Epub 2013 Jul 4.
This study aimed to investigate the role of CD147 in the progression of gastric cancer and the signalling pathway involved in CD147-mediated gastric cancer cell line SGC7901 proliferation and invasion. Short hairpin RNA (shRNA) expression vectors targeting CD147 were constructed to silence CD147, and the expression of CD147 was monitored by quantitative realtime reverse transcriptase polymerase chain reaction and Western blot and further confirmed by immunohistochemistry in vivo. Cell proliferation was determined by Cell Counting Kit-8 assay, the activities of matrix metalloproteinase (MMP)-2 and MMP-9 were determined by gelatin zymography, and the invasion of SGC7901 was determined by invasion assay. The phosphorylation and non-phosphorylation of the mitogen-activated protein kinases, extracellular signal-regulated kinase1/2 (ERK1/2), P38 and c-Jun NH2-terminal kinase were examined by Western blot. Additionally, the ERK1/2 inhibitor U0126 were used to confirm the signalling pathway involved in CD147-mediated SGC7901 progression. The BALB/c nude mice were used to study tumour progression in vivo. The results revealed that CD147 silencing inhibited the proliferation and invasion of SGC7901 cells, and down-regulated the activities of MMP-2 and MMP-9 and the phosphorylation of the ERK1/2 in SGC7901 cells. ERK1/2 inhibitor U0126 decreased the proliferation, and invasion of SGC7901 cells, and down-regulated the MMP-2 and MMP-9 activities. In a nude mouse model of subcutaneous xenografts, the tumour volume was significantly smaller in the SGC7901/shRNA group compared to the SGC7901 and SGC7901/snc-RNA group. Immunohistochemistry analysis showed that CD147 and p-ERK1/2 protein expressions were down-regulated in the SGC7901/shRNA2 group compared to the SGC7901 and SGC7901/snc-RNA group. These results suggest that ERK1/2 pathway involves in CD147-mediated gastric cancer growth and invasion. These findings further highlight the importance of CD147 in cancer progression, indicating that CD147 would be an attractive therapeutic target for gastric cancer.
本研究旨在探讨 CD147 在胃癌进展中的作用，以及 CD147 介导的胃癌细胞系 SGC7901 增殖和侵袭所涉及的信号通路。构建了靶向 CD147 的短发夹 RNA(shRNA)表达载体，沉默 CD147，通过实时定量逆转录聚合酶链反应和 Western blot 监测 CD147 的表达，并在体内进一步通过免疫组化进行确认。通过细胞计数试剂盒-8 测定细胞增殖，通过明胶酶谱法测定基质金属蛋白酶(MMP)-2 和 MMP-9 的活性，通过侵袭测定法测定 SGC7901 的侵袭。通过 Western blot 检测丝裂原活化蛋白激酶，细胞外信号调节激酶 1/2(ERK1/2)、P38 和 c-Jun NH2-末端激酶的磷酸化和非磷酸化。此外，使用 ERK1/2 抑制剂 U0126 来确认 CD147 介导的 SGC7901 进展所涉及的信号通路。使用 BALB/c 裸鼠在体内研究肿瘤进展。结果显示，沉默 CD147 抑制了 SGC7901 细胞的增殖和侵袭，并下调了 SGC7901 细胞中 MMP-2 和 MMP-9 的活性以及 ERK1/2 的磷酸化。ERK1/2 抑制剂 U0126 降低了 SGC7901 细胞的增殖和侵袭，并下调了 MMP-2 和 MMP-9 的活性。在皮下异种移植裸鼠模型中，与 SGC7901 和 SGC7901/snc-RNA 组相比，SGC7901/shRNA 组的肿瘤体积明显较小。免疫组化分析显示，与 SGC7901 和 SGC7901/snc-RNA 组相比，SGC7901/shRNA2 组的 CD147 和 p-ERK1/2 蛋白表达下调。这些结果表明 ERK1/2 通路参与了 CD147 介导的胃癌生长和侵袭。这些发现进一步强调了 CD147 在癌症进展中的重要性，表明 CD147 可能成为胃癌治疗的一个有吸引力的靶点。
