目的:探讨肿瘤相关成纤维细胞( CAFs)对口腔癌细胞增殖、迁移、侵袭的影响,初步分析产生影响的可能原因。方法:体外培养CAFs、舌癌细胞株SCC-9和CAL-27,并建立CAFs与舌癌细胞株共同培养体系;采用红色荧光蛋白标记法观察CAFs对舌癌细胞株增殖能力的影响;利用Transwell小室建立CAFs与舌癌细胞株SCC-9或CAL-27的交互作用模型,观察CAFs对舌癌细胞株迁移和侵袭能力的影响;采用细胞因子芯片检测CAFs单独培养、SCC-9单独培养、CAFs和SCC-9共同培养上清中可溶性细胞因子水平的差异。采用裸鼠成瘤实验观察CAFs对舌癌成瘤能力的影响。结果:培养第5天SCC-9与CAFs共同培养组中SCC-9的数量是初始SCC-9量的(4.41±0.38)倍,明显高于SCC-9单独培养组(3.21±0.35)倍;CAL-27组结果相似。24 h后SSC-9+CAFs组迁移到小室底面的SCC-9细胞与SCC-9单独培养组相比比值为2.6±0.42,CAL-27组比值为3.11±0.46。细胞侵袭研究结果与迁移研究结果类似,共同培养组穿透基质胶的OCC细胞明显增多( P<0.05)。细胞因子抗体芯片分析发现,与细胞单独培养相比,共同培养组培养上清中CCL2、CCL5、IL8水平明显升高。裸鼠成瘤实验结果发现,6周后,SCC-9与CAFs混合注入组的肿瘤体积明显大于SCC-9单独注入组。结论:口腔CAFs能促进舌癌细胞株SCC-9和CAL-27的增殖、迁移和侵袭和肿瘤生长,癌细胞功能的改变可能与微环境中CCL2、CCL5、IL8等细胞因子水平升高有关。%Objective:To elucidate the effects of CAFs on the proliferation,migration and invasion of oral cancer cells and those on tumor growth.Potential mechanism was discussed.Methods:CAFs and SCC-9 or CAL-27 were cocultured.The proliferation of oral cancer cells were detected by measuring the level of RFP.A interaction model between CAFs and SCC-9 or CAL-27 were established with Transwell chambers to abserve the effects of CAFs on the migration and invasion of OCCs.The conditioned medium collected from mono-cultured CAFs or SSC-9 cell,and co-culture were subjected to a cytokine antibody array.Volumes of xenografts were obtained and presented.Results:At day 5,significant difference was found in OCC proliferation between OCC-CAFs group and OCC group.The SCC-9 cell number was up to 4.41±0.38 times as many as the initial number in co-culture group and 3.21±0.35 times in SCC-9 group(P<0.05).The proliferation of CAL-27 was similar to that of SCC-9.SCC-9 cells migrated to the bottom in group SCC-9+CAFs were 2.6± 0.42 times as many as group SCC-9(P<0.05).CAL-27 cells migrated to the bottom in group CAL-27+CAFs were 3.11±0.46 times as many as group CAL-27 ( P<0.05 ) .The similar results were found in invasion analysis.OCCs invaded more quickly towards medium derived from co-cultured cells than that from OCCs alone.Three human cytokines(CCL2,CCL5 and IL-8)were significantly upregulated in conditioned medium from co-cultured cells compared with those from CAFs or SSC-9 alone.Six weeks after injection,we observed that SCC-9 mixed with CAFs generated tumors of greater volume than those generated from SCC-9 cells alone.Conclusion:CAFs Promote the Proliferation,Migration,Invasion of oral cancer cells and tumor growth.Cytokines(CCL2,CCL5 and IL-8)may responsible for the bi-ological function change of cancer cells.
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