Zhang Jing-Cheng, Liu Fan-Rong, Hu Hui-Xian, He Fang, Tu Yan, Wei Bin
Department of Hematology, Jinhua Hospital of Zhejiang University, Jinhua 321000, Zhejiang Province, China.
Department of Patho-logy, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Oct;23(5):1325-30. doi: 10.7534/j.issn.1009-2137.2015.05.019.
To explore whether the chemotactic factor CCL5 is the major factor of diffuse large B cell lymphoma (DLBCL) induced by diabetes or not.
The normal human B cells and DLBCL cells were cultured in vitro; the RT-PCR was used to detect their CCL5 mRNA expression, the human DLBCL cell line and mouse-derived DLBCL cell line A20 were cultured in vitro by using glucose at 5 and 30 mmol/L, respectively, then their CCL5 mRNA expression was detected by PT-PCR; the diabetic mouse model was constructed through peritoneal injection of streptozotocin at low dose in the BALB/c mice; the cell lines with stably high and low expression of CCL5 were established via lentiviral transfection and the cell lines with low and high expression of CCL5 were subcutaneously injected into diabetic mice and mice with normal blood sugar level. According to blood sugar level, the experimental mice were divided into 2 groups: diabetic group (A group) and normal blood sugar group as control (B group); then according to CCL5 expression level, the A group and B group were divided as well into high expression group (A1 group and B1 group) and low expression group (A2 group and B2 group), respectively, the tumor-formation rate, tumor-formation time, tumor size and texture were analyzed, respectively; the CCL5 expression was detected by using HE staining of tumor tissue and immunohistochemical method.
The expression of CCL5 mRNA in human DLBCL cell line cultured in vitro was higher than that in normal B cells (P < 0.05); the expressions of CCL5 mRNA in human DLBCL cells cultured in high sugar concentration in vitro and mouse DLBCL cells were higher than those in cells cultured in low sugar concentration (P < 0.05). The tumor-formation rates in diabetic mice injected with high and low expression of CCL5 cell line A20 were 93.3% in A1 group and 60% in A2 group; the their tumor-formation time was 7.0 ± 0.85 days in A1 group and 9.5 ± 2.8 days in A2 group, while the tumor formation rates in mice with normal blood sugar level were 20% in B1 group and 20% in B2 group, and their tumor-formation time was 12 ± 1.3 days and 14 ± 2.5 days respectively; the CCL5 expression level in tumor tissue of diabetic mice was higher than that in tumor tissue of mice with normal blood sugar level.
The high blood glucose level can casase increase of DLBCL risk and promote the tumor growth; the chemotactic factor CCL5 may play an important role in the growth and migration of DLBCL caused by diabetes mellitus.
探讨趋化因子CCL5是否为糖尿病诱发弥漫性大B细胞淋巴瘤(DLBCL)的主要因素。
体外培养正常人B细胞和DLBCL细胞;采用RT-PCR检测其CCL5 mRNA表达,分别用5 mmol/L和30 mmol/L葡萄糖体外培养人DLBCL细胞系及小鼠来源的DLBCL细胞系A20,然后用PT-PCR检测其CCL5 mRNA表达;通过向BALB/c小鼠腹腔注射低剂量链脲佐菌素构建糖尿病小鼠模型;通过慢病毒转染建立CCL5稳定高表达和低表达的细胞系,将CCL5低表达和高表达的细胞系分别皮下注射到糖尿病小鼠和血糖正常的小鼠体内。根据血糖水平将实验小鼠分为2组:糖尿病组(A组)和血糖正常组作为对照(B组);然后根据CCL5表达水平,A组和B组又分别分为高表达组(A1组和B1组)和低表达组(A2组和B2组),分别分析肿瘤形成率、肿瘤形成时间、肿瘤大小及质地;采用肿瘤组织HE染色及免疫组化方法检测CCL5表达。
体外培养的人DLBCL细胞系中CCL5 mRNA表达高于正常B细胞(P < 0.05);体外高糖浓度培养的人DLBCL细胞及小鼠DLBCL细胞中CCL5 mRNA表达高于低糖浓度培养的细胞(P < 0.05)。注射CCL5高表达和低表达细胞系A20的糖尿病小鼠中,A1组肿瘤形成率为93.3%,A2组为60%;A1组肿瘤形成时间为7.0±0.85天,A2组为9.5±2.8天,而血糖正常小鼠中,B1组和B2组肿瘤形成率均为20%,肿瘤形成时间分别为12±1.3天和14±2.5天;糖尿病小鼠肿瘤组织中CCL5表达水平高于血糖正常小鼠肿瘤组织。
高血糖水平可导致DLBCL风险增加并促进肿瘤生长;趋化因子CCL5可能在糖尿病所致DLBCL的生长和迁移中起重要作用。