Sun Erlin, Fan Xiaodong, Wang Lining, Lei Mingde, Zhou Xiaodong, Liu Chunyu, Lu Bingxin, Nian Xuewu, Sun Yan, Han Ruifa
Department of Urology, Tianjin Institute of Urology, The Second Hospital of Tianjin Medical University, Tianjin 300211, P.R. China.
Department of Gynaecology, Tianjin City Hospital for Gynaecology and Obstetrics, Tianjin 300100, P.R. China.
Oncol Rep. 2015 Jul;34(1):183-94. doi: 10.3892/or.2015.3985. Epub 2015 May 18.
Bacillus Calmette-Guérin (BCG) reduces the recurrence and progression of non-muscle invasive bladder cancer. The present study aimed to investigate the impact of a recombinant hIFN-α2b-secreting BCG (rBCG) on the mouse bladder MB49 cell line and an orthotopic mouse model of bladder cancer. MB49 cells were cultivated in the presence or absence of rBCG, BCG or BCG+hIFN-α2b. Cellular morphology and viability were assessed by microscopy and CCK-8 assay, respectively. Apoptosis was assessed by acridine orange, Hoechst 33258 staining and flow cytometry. MHC-I expression was assessed by flow cytometry. MB49 cells were transplanted into the bladders of C57BL/6 mice administered BCG, rBCG or BCG+hIFN-α2b. Local tissue Fas expression and T cell subsets were assessed by immunohistochemistry. Peripheral blood TNF-α and IL-12 levels were measured by ELISA, and circulating T lymphocyte subsets by flow cytometry. BCG, rBCG and BCG+hIFN-α2b increased the distortion and death of MB49 cells, yet rBCG reduced the proliferation and enhanced apoptosis most substantially. Apoptosis was increased after a 24-h co-culture with rBCG or BCG+hIFN-α2b. Mice administered rBCG survived longer than mice administered BCG (p<0.001), yet this result was not significantly different from mice administered BCG+hIFN-α2b. The average bladder weight was reduced by administration of rBCG (p<0.001). Fas expression and peripheral blood mTNF-α and mIL-12, cell counts of polymorphonuclear leukocytes, monocytes, T lymphocytes and CD4+/CD8+ ratios were significantly increased by all BCG treatments (p≤0.05), yet monocyte and T lymphocyte counts were higher in mice administered rBCG than in mice treated with BCG or BCG+hIFN-α2b (p=0.000). These results indicate that in an orthotopic murine bladder cancer model rBCG possesses superior antitumor activity to BCG+hIFN-α2b.
卡介苗（BCG）可降低非肌层浸润性膀胱癌的复发和进展。本研究旨在探讨分泌重组人干扰素α2b的卡介苗（rBCG）对小鼠膀胱MB49细胞系及膀胱癌原位小鼠模型的影响。将MB49细胞分别在有或无rBCG、BCG或BCG+hIFN-α2b的条件下培养。分别通过显微镜检查和CCK-8法评估细胞形态和活力。通过吖啶橙、Hoechst 33258染色及流式细胞术评估细胞凋亡。通过流式细胞术评估MHC-I表达。将MB49细胞移植到接受BCG、rBCG或BCG+hIFN-α2b处理的C57BL/6小鼠的膀胱中。通过免疫组织化学评估局部组织Fas表达和T细胞亚群。通过ELISA检测外周血TNF-α和IL-12水平，通过流式细胞术检测循环T淋巴细胞亚群。BCG、rBCG和BCG+hIFN-α2b均增加了MB49细胞的变形和死亡，但rBCG最显著地降低了细胞增殖并增强了细胞凋亡。与rBCG或BCG+hIFN-α2b共培养24小时后，细胞凋亡增加。接受rBCG处理的小鼠比接受BCG处理的小鼠存活时间更长（p<0.001），但这一结果与接受BCG+hIFN-α2b处理的小鼠无显著差异。给予rBCG后，膀胱平均重量降低（p<0.001）。所有BCG处理均使Fas表达、外周血mTNF-α和mIL-12、多形核白细胞、单核细胞、T淋巴细胞计数及CD4+/CD8+比值显著增加（p≤0.05），但接受rBCG处理的小鼠的单核细胞和T淋巴细胞计数高于接受BCG或BCG+hIFN-α2b处理的小鼠（p=0.000）。这些结果表明，在原位小鼠膀胱癌模型中，rBCG具有优于BCG+hIFN-α2b的抗肿瘤活性。
