解毒三根汤干预肿瘤相关成纤维细胞及抑制结肠癌侵袭转移的实验研究

目的观察解毒三根汤（虎杖根、水杨梅根、藤梨根）对结肠癌CT-26细胞的迁移能力及对肿瘤相关成纤维细胞（CAFs）转化生长因子β1（transforming growth factor-β1，TGF-β1）、基质金属蛋白酶9（matrix metalloproteinase 9，MMP-9）、α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）的影响。方法 BALB/c小鼠皮下接种CT-26结肠癌细胞（12×106mL）；随机分为3组：正常对照组，模型组，解毒三根汤组。Transwell观察3组血清对CT-26细胞迁移力的影响， ELISA法检测3组CAFs上清液中TGF-β1、MMP-9的表达量，实时荧光定量PCR检测各组CAFs中α-SMA、TGF-β1 mRNA表达量。结果解毒三根汤组透膜结肠癌CT-26细胞数量较模型组明显减少，差异有统计学意义（P<0.01）；24 h及48 h解毒三根汤组刺激的CAFs上清液中TGF-β1、MMP-9的表达量较模型组降低（P<0.05，P<0.01）；解毒三根汤组α-SMA、TGF-β1 mRNA表达量较模型组明显降低，差异有统计学意义（P<0.01）。结论解毒三根汤降低CAFs上清液中TGF-β1、MMP-9的表达量，降低CAFs中α-SMA mRNA、TGF-β1mRNA的表达量，可能是其抑制肿瘤细胞迁移侵袭的机制。

英文摘要:

Objective 〖JP3〗To observe the effect of Jiedu Sangen Decoction （JSD， consisting of Polygonum cuspidatum〖QX）〗〖KG-3〗， Geum Japonicum〖QX）〗 Thumnb， Radix Actinidiae〖QX）〗 Chinensis） on the migration capability of colon cancer CT-26 cells were observed， and on expressions of carcinoma-associated fibroblasts （CAFs） such as transforming growth factor-β1 （TGF-β1）， matrix metalloproteinase 9 （MMP-9）， and α-smooth muscle actin（α-SMA）. Methods The BALB/C mice were subcutaneously inoculated with colon cancer CT-26 cells （1.2×106 mL） and then randomly divided into 3 groups， i.e.， the normal control group， the model group， the JSD treated group. The effects of three different serums on the migration ability of colon cancer CT-26 cells were observed using Transwell. The expression quantities of TGF-β1 and MMP-9 in the supernatant of CAFs were detected using ELISA. The mRNA expression quantities of TGF-β1 and α-SMA in CAFs were detected by real-time fluorescence quantitative PCR. Results The number of semi-permeable film cells in the JSD treated group significantly decreased， when compared with the model group， showing statistical significance （P<0.01）. Compared with the model group， the expressions of TGF-β1 and MMP-9 in the supernatant of CAFs decreased in the JSD treated group at 24 and 48 h， showing statistical difference （P<0.05， P<0.01）. Compared with the model group， the mRNA expressions of TGF-β1 and α-SMA in the JSD treated group obviously decreased， showing statistical difference （P<0.01）. Conclusion JSD could decrease expressions of TGF-β1 and MMP-9 in the supernatant of CAFs， lower mRNA expressions of α-SMA and TGF-β1， which might be possible mechanisms for inhibiting the migration and invasion of tumor cells.

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