槐耳浸膏对人胃癌MKN-45、SGC-7901细胞凋亡及基质金属蛋白酶2、基质金属蛋白酶9表达的影响  

Effect of trametes robiniophila on the apoptosis of human gastric carcinoma MKN-45 and SGC-7901 cells and the expressions of matrix metalloproteinase 2 and matrix metalloproteinase 9

作　　者：高运宾 李晓文 全秀莲[2] 吕金燕[2] 冀学宁[2] Gao Yunbin;Li Xiaowen;Quan Xiulian;Lyu Jinyan;Ji Xuening(Department of Oncology, Jining No.1 People's Hospital, Jining 272011, China;Department of Oncology, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China)

机构地区：[1]济宁市第一人民医院肿瘤科,272011 [2]大连大学附属中山医院肿瘤科,116001

出　　处：《肿瘤研究与临床》2019年第4期217-222,共6页Cancer Research and Clinic

摘　　要：目的分析槐耳浸膏对人胃癌低分化细胞株MKN-45和中分化细胞株SGC-7901细胞凋亡及对细胞侵袭、转移相关的基质金属蛋白酶2(MMP-2)和MMP-9表达的影响。方法分别用0(阴性对照组)、5、10、20 mg/ml槐耳浸膏作用于MKN-45和SGC-7901细胞24 h,经吖啶橙-溴化乙啶(AO-EB)双荧光染色后,显微镜下观察其形态学表现;作用24、48、72 h后,采用流式细胞术检测细胞凋亡率;作用24 h后,采用反转录聚合酶链反应(RT-PCR)检测MMP-2、MMP-9 mRNA表达情况,以凝胶电泳后条带吸光度值为其表达量。结果荧光显微镜下观察发现,阴性对照组细胞绿染(正常细胞),大小均一,结构完整;随着槐耳浸膏浓度增加,凋亡细胞和坏死细胞比例增加,其中凋亡细胞黄染,形态不规则,细胞膜可见芽状突起,细胞核固缩、碎裂,染色质浓染并沿核膜分布;坏死细胞染成红色,大小、形态较均一。槐耳浸膏作用于MKN-45细胞24 h后,阴性对照组和5、10、20 mg/ml槐耳浸膏组的细胞凋亡率分别为(6.5±0.8)%、(14.6±1.0)%、(18.0±1.1)%、(23.1±1.2)%(F=333.972,P<0.01),作用48 h后分别为(7.3±1.2)%、(18.3±1.6)%、(24.5±1.3)%、(27.2±1.7)%(F=528.432,P=0.001),作用72 h后分别为(7.5±0.9)%、(50.2±1.6)%、(58.0±1.9)%、(69.0±1.4)%(F= 3 814.238,P<0.01)。槐耳浸膏作用于SGC-7901细胞24 h后,阴性对照组和5、10、20 mg/ml槐耳浸膏组细胞凋亡率分别为(12.9±1.0)%、(19.4±1.2)%、(22.0±1.7)%、(23.0±1.9)%(F=120.190,P<0.01),作用48 h后分别为(10.2±1.3)%、(40.9±1.4)%、(51.6±1.9)%、(66.2±1.9)%(F=1 281.342,P<0.01),作用72 h后分别为(27.4±1.8)%、(49.7±1.4)%、(65.1±1.4)%、(69.0±2.0)%(F=1 112.767,P<0.01)。槐耳浸膏诱导两种细胞凋亡时表现为时间和剂量依赖性(均P<0.01)。相同作用条件,SGC-7901细胞凋亡率较MKN-45细胞有增高趋势。槐耳浸膏作用于胃癌MKN-45细胞后,阴性对照组和5、10、20 mg/ml槐耳浸膏组MMP-2 mRNA表达量(吸光度)分别为0.64±0.02、0.4Objective To analyze the effect of trametes robiniophila on the apoptosis and the expressions of invasion and metastasis related matrix metalloproteinase 2 (MMP-2) and MMP-9 in human gastric carcinoma poorly differentiated cell line MKN-45 and medium differentiated cell line SGC-7901. Methods Human gastric carcinoma cell lines MKN-45 and SGC-7901 incubated with trametes robiniophila at the different concentrations [0 (the negative control group), 5, 10, and 20 mg/ml] for 24 h. When bifluorescently stained with acridine orange-ethidium bromide (AO-EB), morphology was observed by using microscopy. Flow cytometry was used to test the apoptosis ratio after 24, 48, and 72 h. Reverse transcription polymerase chain reaction (RT-PCR) was applied to analyze the expressions of mRNA of MMP-2 and MMP-9 after 24 h, and the absorbance values of the bands after gel electrophoresis were used as their expression levels. Results Under the fluorescence microscope, the cells of the negative control group were green (normal cells), and the membrane was even in size and integrity. The proportion of apoptotic and necrotic cells was increased with the concentration enrichment of trametes robiniophila. The apoptotic cells were yellow, and their cell membrane lost integrity. Apoptotic bodies were found in cancer cells, and cell membrane showed bud projection. The nuclei of the apoptotic cells showed brightly condensed chromatin or fragmented. Chromatin was strongly stained and located in karyotheca. The necrotic cells were dyed red with integrity of size. The apoptotic ratio of MKN-45 cell line induced by trametes robiniophila after 24 h was (6.5±0.8)%,(14.6±1.0)%,(18.0±1.1)%, and (23.1±1.2)%, respectively (F = 333.972, P < 0.01) in negative control group and 5, 10, and 20 mg/ml trametes robiniophila group. After 48 h, the apoptotic ratio was (7.3±1.2)%,(18.3±1.6)%,(24.5±1.3)%, and (27.2±1.7)%, respectively (F = 528.432, P=0.001);after 72 h, the apoptotic ratio was (7.5±0.9)%,(50.2±1.6)%,(58.0±1.9)%, and (69.0±1.4)%, respectiv

关 键 词：胃肿瘤 细胞凋亡 肿瘤侵润 肿瘤转移 槐耳浸膏 

分 类 号：R285.5[医药卫生—中药学]