法氏囊活性肽-Ⅱ对肿瘤细胞增殖的抑制作用及其可能机制  

Inhibitory effect and mechanism of Bursal-derived pentapeptide-Ⅱ on tumor cell proliferation and its possible medanism

作　　者：郭香玲[1] 王臣[1] 李小康[1] 汪洋[1] 吴庭才[1] 陈溥言[2] 

机构地区：[1]河南科技大学兽医肿瘤免疫学重点实验室,河南洛阳471003 [2]南京农业大学农业部动物细菌学重点实验室,江苏南京210095

出　　处：《中国肿瘤生物治疗杂志》2015年第1期22-27,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学青年基金资助项目(No.31101792;No.31201928);河南省高校青年骨干教师资助项目(No.2012GGJS-07)~~

摘　　要：目的:研究法氏囊活性肽(Bursal-derived pentapeptide,BPP)-Ⅱ对肿瘤细胞增殖的抑制作用及其可能的作用机制。方法:采用不同质量浓度(0.02、0.2、2、20μg/ml)的BPP-Ⅱ分别处理小鼠B细胞淋巴瘤细胞WEHI-231、人鼻咽癌细胞CNE、大鼠肝癌细胞RH-35和正常细胞系人胚肾细胞293、猪肾细胞PK15、中国仓鼠卵巢细胞CHO,48 h后采用MTT法检测细胞增殖情况;以双荧光素酶报告系统和Western blotting方法检测BPP-Ⅱ对荧光素酶标记的p53-Luc质粒转染的Vero细胞中p53转录活性和P53蛋白表达的影响;利用噬菌体随机12肽库筛选BPP-Ⅱ特异性结合肽,人工合成BPP-Ⅱ结合肽,采用MTT法检测BPP-Ⅱ结合肽对BPP-Ⅱ抗WEHI-231细胞增殖能力的影响。结果:2和20μg/ml的BPP-Ⅱ对肿瘤细胞WEHI-231、CNE、RH-35的增殖均有抑制作用(均P<0.05),而对正常细胞系293、PK15、CHO的增殖均不表现出抑制作用。BPP-Ⅱ明显激活p53基因的转录,并上调P53蛋白的表达。应用噬菌体展示技术筛选获得3个BPP-Ⅱ结合肽P3-12、P5-12、P6-12,其中2和20μg/ml的P3-12能显著抑制BPP-Ⅱ的抗WEHI-231细胞增殖能力[(97.5±3.4)%、(98.9±3.5)%vs(86.3±1.9)%,P<0.05];20μg/ml的P5-12和P6-12均能显著抑制BPP-Ⅱ的抗WEHI-231细胞增殖能力[(96.7±3.1)%、(95.4±3.8)%vs(86.3±1.9)%,P<0.05]。结论:BPP-Ⅱ可特异性抑制WEHI-231、CNE、RH-35等肿瘤细胞增殖,其机制可能与激动p53信号通路有关。Objective: To study the anti-tumor activity and mechanism of Bursal-derived pentapeptide-Ⅱ( BPP-Ⅱ).Methods: Tumor cells including murine B-cell lymphoma WEHI-231 cells,human nasopharyngeal carcinoma CNE cells and rat hepatoma RH-35 cells and normal cells including human embryonic kidney 293 cells,pig kidney PK15 cells and Chinese hamster ovary CHO cells were stimulated with BPP-Ⅱ at 0. 02 μg / ml,0. 2 μg / ml,2 μg / ml and 20 μg / ml for 48 h. Cell viability was then measured by MTT assays,p53 luciferase activity and the expression of P53 at the protein level were assessed,and the effect of BPP-Ⅱ binding peptides biopaned from a phage display 12-mer random peptide library on BPP-Ⅱ-mediated inhibition of WEHI-231 cell proliferation was determined by MTT assays. Results: BPP-Ⅱ inhibited the proliferation of tumor cells but not normal cells,activated p53 transcription,and increased P53 protein content. After four rounds of biopanning,three BPP-Ⅱ binding peptides were obtained: QSLPSPLWIQQS( P3-12),DRMPDSAWTTRK( P5-12) and ALWPPNLHAWVP( P6-12). P3-12 significantly inhibited the anti-proliferative activity of BPP-Ⅱ at both 2μg /ml( 97. 5 ± 3. 4) % and 20 μg /ml( 98. 9 ± 3. 5) % as compared with the control( 86. 3 ± 1. 9) % in WEHI-231 cells( P < 0. 05). At 20 μg/ml,both P5-12( 96. 7 ± 3. 1) % and P6-12( 95. 4 ± 3. 8) % inhibited the anti-proliferative activity of BPP-Ⅱ as compared with the control( 86. 3 ± 1. 9%) in WEHI-231 cells( P < 0. 05). Conclusions: BPP-Ⅱ appears to have an antitumor activity,possibly attributable to p53 activation. Taken together,our findings suggest a significant clinical importance for BPP-Ⅱ from the perspective of cancer therapy.

关 键 词：法氏囊活性肽-Ⅱ 肿瘤细胞 抗肿瘤 结合肽 p53信号通路 

分 类 号：R730.2[医药卫生—肿瘤]