下调PBLD的表达对胃癌增殖、侵袭的影响

The effects of reduced PBLD expression on proliferation and migration of gastric cancer

目的检测吩嗪生物合成类结构域蛋白(PBLD)在胃癌与癌旁正常组织中的表达,探讨其表达下调对胃癌细胞增殖、侵袭和凋亡的影响。方法采集胃癌220例、癌旁正常组织79例及患者临床信息,采用免疫组化技术检测PBLD的表达,分析PBLD的水平与患者年龄、性别、肿瘤分期和转移等临床特点的关系。同时构建PBLD干扰表达载体,在胃癌细胞AGS中干扰PBLD的表达,通过RT-PCR鉴定干扰效果,通过CCK-8实验检测细胞增殖能力,通过Transwell实验检测细胞侵袭能力,通过Annxin V-PI染色法流式细胞计数检测细胞的凋亡水平。结果PBLD在胃癌组织中的表达阳性率为27.7%,较癌旁正常组织低,癌旁组织阳性率为63.3%(P<0.001)。PBLD与患者的TNM分期(P=0.001)、淋巴结转移(P=0.016)和分化(P=0.001)相关。干扰PBLD的表达,胃癌细胞AGS的增殖能力增强(P<0.01),Transwell实验结果提示,干扰组穿过基质膜的细胞数多于未干扰组(P<0.05),PBLD干扰后侵袭能力增强。干扰PBLD组总凋亡率为(7.36±0.78)%,较空载体组总凋亡率(11.76±0.64)%降低(P=0.007),干扰PBLD组晚期调亡率为(5.56±1.7)%,较空载体组晚期调亡率(10.75±1.02)%降低(P=0.015),但早期凋亡率无统计学意义。结论PBLD在胃癌组织中表达水平降低,这与胃癌细胞的增殖、侵袭能力和晚期凋亡相关,PBLD是胃癌发生发展中的候选抑癌基因。

Objective To analyze the effects of down-regulation of phenazine biosynthesis-like domain-containing protein(PBLD) on proliferation,migration and apoptosis of gastric cancer(GC) cells by detecting the expression level of PBLD in the GC and para-carcinoma normal tissues. Methods Collected 220 GC tumor tissues and 79 para-carcinoma normal gastric tissues. Recorded the clinic information. Used immunohistochemistry method to detect expression level of PBLD in the tissues. Analyzed the correlation of PBLD and clinic characteristics include the age,gender,tumor stage,and migration. Constructed the plasmids for interference of the PBLD expression. Then,used the plasmids to down regulate the expression of PBLD in AGS cells. The cells transfected with empty plasmids were used as the control. Detected the interference efficiency by RT-PCR. Detected the cell proliferation ability by CCK-8 method and migration ability by Transwell assay. Detected the rate of apoptosis using Annexin V-PI and Flow cytometry. Results The positive rate of PBLD in GC tissues was 27. 7%. It was lower than that in para-carcinoma normal tissues( 63. 3%)( P < 0. 001). The expression of PBLD was related with TNM stage( P = 0. 001),lymphatic metastasis( P = 0. 016) and differentiation( P = 0. 001). After interference expression of PBLD,the proliferation ability of AGS cells was increased( P < 0. 01). In Transwell assay,the cells that transfer to the basilar membrane in test group were more than those of control group. So the migration ability after interference expression of PBLD was promoted( P < 0. 05). The total apoptosis rate of interference group was( 7. 36 ± 0. 78) %,which was lower than that of control group( 11. 76 ± 0. 64) %( P = 0. 007). The late apoptosis rate( 5. 56 ± 1. 7) % of interference group was also lower than that of control group( 10. 75 ± 1. 02) %( P = 0. 015). But the early apoptosis rate had no statistical significance. Conclusion PBLD has lower expression level in GC tissues. The down regulation of PBLD is related with the proliferation,migration and late apoptosis rate of GC cells. PBLD is a candidate cancer suppressor gene for GC.