摘要
目的探究卡介苗对膀胱肿瘤细胞及代谢产物的作用和影响,对卡介苗治疗膀胱肿瘤细胞的可能机制进行初步的探索。方法通过膀胱灌注N-甲基亚硝基脲(MNU)诱导并建立大鼠的膀胱肿瘤模型,将膀胱肿瘤细胞和正常移行上皮细胞进行原代培养。按照培养液的不同成分将其分为5组,利用酶联免疫吸附试验(ELISA)法检测各组细胞上清液中的肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)的浓度。利用噻唑蓝(MTT)法测定卡介苗抑制肿瘤细胞生长的药物浓度。利用末端脱氧核苷酸转移酶介导缺口末端标记法(TUNEL)检测细胞凋亡情况。结果在灌注的15只大鼠中,2只大鼠在灌注2次后死亡,3只大鼠灌注3次后死亡,解剖死亡大鼠均未发现明显的肿瘤。再对其余10只大鼠完成膀胱灌注后,其中8只大鼠解剖后发现有明显的肿瘤。MTT结果显示:卡介苗对膀胱肿瘤细胞的生长具有抑制作用,其抑制率与卡介苗的浓度呈现正相关。ELISA检测各组细胞上清液中TNF-α浓度在B组为(160.654±5.775)ng/L,D组为(124.443±4.972)ng/L的浓度,与其他三组相比差异有显著性;IL-10浓度在B组为(16.973±3.428)ng/L,E组为(20.327±2.721)ng/L,蛋白水平明显高于其他三组。各组细胞均未发现细胞凋亡。对原代膀胱肿瘤细胞的鉴定显示:利用HE染色可发现肿瘤细胞的细胞核体积增大,核浆比增高,有的可发现细胞核仁数目增加,有的细胞核呈现墨水滴样,核仁比较明显。结论卡介苗对大鼠膀胱肿瘤细胞的增长具有抑制作用,没有发生细胞凋亡,且肿瘤细胞自身分泌的IL-10、TNF-α可以参与调节。
ObjectiveTo investigate the effect of bacillus Calmette Guérin(BCG)on bladder cancer cells and their metabolites,and to preliminarily explore the possible mechanisms of BCG in the treatment of bladder cancer.MethodsThe rat model of bladder cancer was induced by intravesical instillation with N methylnitrosourea(MNU).Bladder cancer cells and normal transitional epithelial cells were isolated and primarily cultured,and were divided into5groups according to the different components of the culture medium.The concentration of tumor necrosis factorα(TNFα)and interleukin10(IL10)in the supernatant of each group was detected by enzyme linked immunosorbent assay(ELISA).The concentration of BCG to inhibit the cancer cell growth was determined by MTT assay.Apoptosis of bladder cancer cells was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).ResultsAmong the15rats,2rats died after2times of instillation,and3rats died after3times of instillation,without obvious tumors found at autopsy.The other10rats were killed after completion of the intravesically instillation of MNU,and obvious tumors were found in8of them after dissection.The results of MTT assay showed that BCG had an inhibitory effect on the growth of bladder cancer cells,and the inhibitory rate was positively correlated with the concentration of BCG.The results of ELISA showed that the concentrations of TNFαin the supernatant of groups B and D were(160654±5775)ng/L and(124443±4972)ng/L,respectively,with significant differences from those of the other three groups.The concentrations of IL10in the groups B and E were(16973±3428)ng/L and(20327±2721)ng/L,significantly higher than those of the other three groups.Apoptosis of cancer cells was not found in all groups.HE staining of the primary bladder cancer cells showed that the volume of cell nucleus was increased,and the nucleo cytoplasmic ratio was increased.The number of nucleoli in some cells was increased and some nuclei appeared like ink drops with prominent nucleoli.Con
作者
吴金生
郑传秋
王清明
纪萌
孙立江
WU Jin sheng;ZHENG Chuan qiu;WANG Qing ming;JI Meng;SUN Li jiang(Qingdao University, Qingdao 266071, China;Weifang Medical University, Weifang 261053, China)
出处
《中国比较医学杂志》
北大核心
2017年第11期56-59,74,共5页
Chinese Journal of Comparative Medicine
