胃癌BCG-823细胞对大鼠骨髓间充质干细胞CD34、CD44表达的影响

Establishment and observation of rat model of gastric cancer BCG-823 cells on the expression of CD34 and CD44 in bone marrow mesenchymal stem cells

目的探究人胃癌BCG-823细胞对大鼠骨髓间充质干细胞(BMSCs)模型CD34、CD44表达的影响。方法分离并培养健康大鼠,对照组直接进行常规BMSCs培养,实验组给予人胃癌BCG-823细胞共培养,培养周期均为7 d。检测各组BMSCs的细胞增殖率、生长形态、细胞分裂周期分布及CD34、CD44表达水平,并比较两组培养BMSCs期间各指标的差异。结果对照组BMSCs贴壁生长并铺满培养瓶底,细胞呈长梭形,镜下观察细胞轮廓不清,具有较强的折光性;观察组BMSCs细胞团状生长并散在分布,细胞形态不规则,与人胃癌BCG-823细胞形态相似。培养7 d后行流行细胞术(FCM)检测细胞分裂周期,观察组BMSCs G1周期占比明显低于对照组,S周期占比及G2/M占比均明显高于对照组(P<0.01)。两组BMSCs在培养1 d时细胞增殖率差异无统计学意义(P>0.05);培养2~7 d时,观察组BMSCs增殖率明显高于对照组(P<0.01)。培养7 d后,观察组CD34阳性表达率明显高于对照组,CD44阳性表达率明显低于对照组(P<0.05)。结论人胃癌BCG-823细胞对大鼠BMSCs增殖速度及CD34、CD44表达具有显著影响,可诱导大鼠BMSCs向人胃癌BCG-823细胞分化。

Objective To establish and observe the rat model of gastric cancer BCG-823 cells on the expression of CD34 and CD44 in bone marrow mesenchymal stem cells（ BMSCs）.Methods Six healthy male Sprague-Dawley rats were sacrificed by cervical dislocation,and the femurs and tibia were isolated and cultured. BMSCs were isolated and cultured. The control group was cultured with normal BMSCs.The observation group was co-cultured with human gastric cancer BCG-823 cells. The culture period was 7 days. The proliferation rate,growth morphology,cell division cycle distribution and CD34 and CD44 expression levels were detected and compared. Results BMSCs in the control group were adherent and adhered to the bottom of the culture. The morphology of the cells was long spindle,and the cells were unclear and had a strong refraction. BMSCs in the observation group grew and scattered,and the cell morphology was irregular and similar to that of human gastric cancer BCG-823 cells. The cell cycle of FCM was significantly lower than that of control group（ P〈0.05）. The ratio of G1 phage of the BMSCs in the observation group was significantly lower than that in control group（ P〈0.05）. The ratio of S-cycle and G2/M were significantly higher than that of control group（ P〈0.01）. There was no significant difference in the proliferation rate of BMSCs between the two groups on the first day of culture（ P〈0.05）. The proliferation rate of BMSCs in observation group was significantly higher than that in control group at the 2 ~ 7 days of culture（ P〈0.01）. The positive expression rate of CD34 in observation group was significantly higher than that in control group（ P〈0.05）. The positive rate of CD44 in observation group was significantly lower than that in control group（ P〈0.05）.Conclusions Human gastric cancer BCG-823 cells has a significant effect on the proliferation rate and CD34 and CD44 expression of BMSCs from rat,and could induce differentiation of rat BMSCs into human gastric cancer BCG-823 cells.