CD14在胃癌中的表达及其阳性亚群增殖和侵袭特性

Expression of CD14 in gastric cancer and proliferative and invasive characteristics of positive subpopulations

目的 探讨CD14在胃癌中的表达及其阳性亚群的体外增殖和侵袭特性。方法 通过免疫荧光双染法检测已经被肿瘤干细胞标志物CD133标记的人胃癌和其癌旁组织石蜡切片中CD14的表达,定位和定性CD14。以CD14为标志物,原代培养人胃癌细胞,以免疫磁珠法分选,分别获得CD14^(+)胃癌细胞和CD14-胃癌细胞,实验分为CD14^(+)胃癌细胞组和CD14-胃癌细胞组;用噻唑蓝比色法检测2组细胞增殖能力,用Transwell实验检测2组细胞侵袭能力,用蛋白质印迹法检测2组细胞Toll样受体4(TLR4)蛋白的表达水平。结果 标记CD133的胃癌肿瘤干细胞同时表达CD14,胃癌组织和癌旁组织CD14、CD133和CD14/CD133共表达阳性率分别为(19.46±2.58)%、(25.15±3.07)%、(13.75±0.91)%;癌旁组织CD14、CD133和CD14/CD133共表达阳性率分别为(1.12±0.24)%、(1.53±0.03)%、(0.95±0.11)%,2组比较,差异均有统计学意义(均P<0.01)。CD14^(+)胃癌细胞组和CD14-胃癌细胞组增殖能力分别为0.56±0.09和0.35±0.02,穿膜细胞数分别为57.51±1.36和38.27±2.09,TLR4表达水平分别为0.64±0.13和0.21±0.05,2组细胞比较,差异均有统计学意义(均P<0.01)。结论 CD14可作为胃癌肿瘤干细胞相对特异性的表面标志物,CD14^(+)胃癌细胞亚群具有一定的肿瘤干细胞特性,并有较强的体外增殖和侵袭能力,其机制可能与上调TLR4蛋白表达水平相关。

Objective To investigate the expression of CD14 in gastric cancer and the proliferation and invasion characteristics of positive subgroups in vitro.Methods CD14 expression,localization and characterization of CD14 in paraffin sections of human gastric cancer and adjacent tissues labeled with tumor stem cell marker CD133 were detected by immunofluorescence double staining.U tsing CD14asa marker,the original cultured human gastric cancer cells were sorted by immunomagnetic beads to obtain CD14^(+)gastric cancer cells and CD14-gastric cancer cells,respectively.The experiment was divided into CD14 gastric cancer cell group and CD14^(+)gastric cancer cell group.The proliferative ability of the two groups was detected by thiazole blue colorimetry,the invasion ability of the two groups were detected by Transwell assay,and the expression level of Toll-like receptor 4(TLR4)protein in the two groups were detected by Western blotting.Results The CD133-labeled gastric cancer stem cells expressed CD14 at the same time,and the positive rates of CD14,CD133 and CD14/CD133 in gastric and paracancer tissues were(19.46±2.58)%,(25.15±3.07)%,(13.75±0.91)%,which in paracancer tissues were(1.12±0.24)%,(1.53±0.03)%,(0.95±0.11)%,the difference between the two groups was statistically significant(all P<0.01).The proliferation capacity of CD14^(+)gastric cancer cell group and CD14~gastric cancer cell group were 0.56±0.09 and 0.35±0.02,the number of perforated cells were 57.51±1.36 and 38.27±2.09,and the expression level of TLR4 were 0.64±0.13 and 0.21±0.05,respectively,the differences were all statistically significant(all P<0.01).Conclusion CD14 can be used as a relatively specific surface marker of gastric cancer stem cells.CD14*gastric cancer cell subpopulation has certain characteristics of tumor stem cells,and has strong ability of proliferation and invasion in vitro.The mechanism may be related to the up-regulation of TLR4 protein expression level.