人胃癌组织中间充质干细胞对胃癌荷瘤鼠瘤体生长的影响

Effect of mesenchymal stem cells from human gastric cancer on the tumor growth of gastric cancer-bearing mice

目的 观察人胃癌组织来源的间充质干细胞（hGC-MSCs）对胃癌荷瘤鼠瘤体生长的影响.方法 培养胃癌细胞株SGC-7901,将浓度为1×10^7个/只的细胞接种于BABL/c裸鼠右侧背部皮下,建立荷瘤鼠模型.经不同剂量（0.5×10^6、1.0 ×10^6、2.0×10^6个/只）的hGC-MSCs作用于荷瘤鼠为实验组,磷酸盐缓冲液（PBS）作用于荷瘤鼠为对照组.测量肿瘤体积变化,采用免疫组织化学染色、实时定量聚合酶链反应（Real-time PCR）、Western blot检测增殖细胞核抗原（PCNA）和基质金属蛋白酶-9 （MMP-9）在各组中的表达.结果 对照组与各实验组的肿瘤体积分别为（1.06±0.15）、（2.21 ±0.44）、（3.38±1.23）、（4.60±0.86） cm3,各组差异有统计学意义（P＜0.05）.免疫组织化学结果示PCNA在对照组和各实验组中的吸光度（A）值分别为0.349±0.008、0.376±0.013、0.441 ±0.012、0.500±0.022;MMP-9在4组中的A值分别为0.341±0.027、0.429±0.015、0.474±0.006、0.527±0.033,各组之间差异有统计学意义（P＜0.01）.Real-time PCR证实hGC-MSCs能够上调PCNA mRNA的表达,在对照组和各实验组分别为1.000±0.000、1.640±0.013、1.890±0.021、2.150±0.021;MMP-9 mRNA在4组中的表达分别为1.000±0.000、1.810±0.101、2.310±0.004、2.700±0.034,各组之间差异有统计学意义（P＜0.05）.Western blot检测对照组和各实验组PCNA的表达分别为14.99 ±0.90、17.64 ±0.90、25.11 ±0.40、28.31±1.10;MMP-9在4组中的表达分别为18.48±1.30、32.95±1.20、67.26±1.20、141.16±3.80,各组之间差异有统计学意义（P＜0.01）.结论 hGC-MSCs在体内能促进胃癌荷瘤鼠瘤体的生长,其机制可能与上调PCNA和MMP-9的表达有关.

Objective To investigate the effect of mesenchymal stem cells from human gastric cancer （hGC-MSCs） on the tumor growth of gastric cancer-bearing mice.Methods Gastric cancer cell line SGC-7901 cells were cultured and inoculated subcutaneously in the right back of BABL/c nude mice with a number concentration of 1 × 107 for each one to form the mice tumor-bearing model.The tumor-bearing mice receiving different doses （0.5 × 106,1.0 × 106,2.0 × 106） of hGC-MSCs were classified into the experimental groups,while those receiving PBS served as the control group.The changes in tumor volume were measured,and the expression of proliferating cell nuclear antigen （PCNA） and matrix metalloproteinase （MMP）-9 was detected by immunohistochemistry,real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting in each group.Results The tumor volume in the control group and the experimental groups was （1.06 ± 0.15）,（2.21 ± 0.44）,（3.38 ± 1.23）,and （4.60 ± 0.86） cm3,respectively （P 〈 0.05）.The immunohistochemical results showed that the average optical density of PCNA in the control group and experimental groups was 0.349 ± 0.008,0.376 ± 0.013,0.441 ±0.012 and 0.500 ±0.022,respectively,and that of MMP-9 was 0.341 ±0.027,0.429 ± 0.015,0.474 ± 0.006 and 0.527 ± 0.033,respectively （P 〈 0.01）.The Real-time PCR confirmed that the mRNA expression of PCNA in the control group and experimental groups were 1.000 ± 0.000,1.640 ± 0.013,1.890 ±0.021 and 2.150 ±0.021,respectively,and that of MMP-9 was 1.000 ±0.000,1.810 ± 0.101,2.310 ± 0.004 and 2.700 ± 0.034 respectively （P 〈 0.05）.Western blotting revealed the expression of PCNA in the control group and the experimental groups was 14.99 ±0.90,17.64 ±0.90,25.11 ± 0.40 and 28.31 ± 1.10,respectively,and that of M MP-9 was 18.48 ± 1.30,32.95 ± 1.20,67.26 ± 1.20 and 141.16 ± 3.80,respectively （P 〈 0.01）.Conclusion hGC-MSCs can promote the tumor growth of gastric cancer-bearing mice in vivo,and its mechanism may be associated with the upregulated expression of PCNA and MMP-9.