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、20 μg/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20 μg/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20 μg/ml的P3-12能显著抑制BPP-Ⅱ的抗WEHI-231细胞增殖能力\[（97.5±3.4）%、（98.9±3.5）% vs （86.3±1.9）%， P<0.05 \]；20 μg/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。

Objectiv : 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 2 μg/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.

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）。