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目的:研究口腔癌成纤维细胞对口腔癌细胞MMP-2、9表达的影响。方法:采用明胶酶谱法和免疫细胞化学法分析MMP-2、MMP-9在口腔癌细胞与口腔癌成纤维细胞(CAFs)中共培养前后表达的变化。结果:①单独培养的Tca-8113细胞中未检测到MMP-2、9表达,单独培养的CAFs中只检测到微弱活性的MMP-2,未检测到活性形式的MMP-9。直接共培养后,上清液中MMP-9的活性随CAFs细胞数量的增加而增加,各条带的平均积分灰度值分别为82.15±0.45、96.40±0.32、103.87±0.47,各组间比较有显著性差异(P<0.05)。而MMP-2活性略有升高。间接共培养后,MMP-2的活性随CAFs细胞数量的增加而增加,各条带的平均积分灰度值分别为84.15±0.25、94.40±0.38、105.87±0.57,各组间比较有显著性差异(P<0.05)。未检测到活性形式的MMP-9。②免疫组化结果表明直接共培养(1∶3比例)后,Tca-8113表达MMP-9较对照组增强,尤其在被CAFs环绕的Tca-8113表达更强。间接共培养(1∶3比例)后,Tca-8113表达MMP-2也较对照组增强。结论:MMP-2表达增加可能受到源自口腔癌相关成纤维细胞可溶性因子的旁分泌介导,而MMP-9表达可能需要口腔癌细胞与癌相关成纤维细胞的直接接触。
Objective: To study the effect of oral cancer fibroblasts on the expression of MMP-2 and 9 in oral cancer cells. Methods: The changes of MMP-2 and MMP-9 expression in oral cancer cells and oral cancer fibroblasts (CAFs) before and after co-culture were analyzed by gelatin zymography and immunocytochemistry. Results: ①The expression of MMP-2 and MMP-9 was not detected in cultured Tca-8113 cells. Only weakly active MMP-2 was detected in CAFs cultured alone, and no active form of MMP-9 was detected. After direct co-culture, the activity of MMP-9 in the supernatant increased with the increase of the number of CAFs cells. The average integral gray value of each band was 82.15 ± 0.45, 96.40 ± 0.32 and 103.87 ± 0.47, respectively There was significant difference (P <0.05). While MMP-2 activity slightly increased. After indirect co-culture, the activity of MMP-2 increased with the increase of the number of CAFs cells, the average integral gray value of each band was 84.15 ± 0.25,94.40 ± 0.38,105.87 ± 0.57, there was a significant difference between the groups (P <0.05). No active form of MMP-9 was detected. ② Immunohistochemistry results showed that after direct co-culture (1: 3 ratio), Tca-8113 expression of MMP-9 increased compared with the control group, especially in Tca-8113 surrounded by CAFs. Indirect co-culture (1: 3 ratio), Tca-8113 expression of MMP-2 also increased compared with the control group. CONCLUSION: Increased MMP-2 expression may be mediated by paracrine origin of soluble factors from oral cancer-associated fibroblasts, whereas MMP-9 expression may require direct contact between oral cancer cells and cancer-associated fibroblasts.