抑制皮肤鳞癌细胞增殖的PEDF短肽筛选及其透皮性分析  

Screening of PEDF short peptides for inhibiting proliferation of skin squamous cell carcinoma cells and analysis of their transdermal properties

作　　者：李春明[1] 刘藕根[1] 彭亚婷 张淑兰[1] 张颖鹏[1] 刘志刚[1] LI Chunming;LIU Ougen;PENG Yating;ZHANG Shulan;ZHANG Yingpeng;LIU Zhigang(The Second Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第二附属医院,南昌330006

出　　处：《山东医药》2018年第32期21-24,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81260238);江西省青年科学基金资助项目(20132BAB215021)

摘　　要：目的筛选抑制皮肤鳞癌细胞增殖的6~7氨基酸(aa)的色素上皮衍生因子(PEDF)短肽,并检测其透皮性。方法将PEDF长肽(301~418 aa)分成6段,人工合成相对应的6条18~20 aa短肽,记为P1~P6,采用CCK-8法筛选出可抑制皮肤鳞癌SCL-1细胞增殖的18~20 aa短肽(短肽浓度均为100 ng/mL)。将上述筛选出的18~20 aa短肽均分成3段,人工合成与其对应的6~7 aa短肽,参照上法筛选出可抑制皮肤鳞癌SCL-1细胞增殖的6~7 aa短肽(短肽浓度均为100 ng/mL)。生物素标记筛选出的6~7 aa短肽,将其外涂于裸鼠的局部皮肤,2 h后分离局部皮肤组织,采用免疫荧光法观察其皮肤渗透性。结果 18~20 aa短肽P1、P2均可抑制皮肤鳞癌SCL-1细胞增殖(P均<0.05),P3~P6对皮肤鳞癌SCL-1细胞增殖均无明显影响(P均>0.05)。P1、P2对应的6~7 aa短肽P1-1、P1-2、P2-2均可抑制皮肤鳞癌SCL-1细胞增殖(P均<0.05),P1-3、P2-1、P2-3对皮肤鳞癌SCL-1细胞增殖均无明显影响(P均>0.05)。经P1-1、P1-2、P2-2短肽外涂的裸鼠皮肤组织血管内及血管壁均可检测到红色荧光。结论成功筛选出可抑制皮肤鳞癌细胞增殖的6~7 aa PEDF短肽P1-1、P1-2、P2-2,其透皮性均较好,有望应用于皮肤鳞癌的治疗。Objective To screen out the 6-7 amino acid(aa)pigment epithelium derived factor(PEDF)short peptides which can inhibit the proliferation of skin squamous cell carcinoma cells and to detect their transdermal properties.Methods The PEDF long peptide(301-418 aa)was divided into 6 segments,and 6 short peptides(18-20 aa)were synthesized,which were recorded as P1 to P6.CCK-8 was used to screen out the 18-20 aa short peptide that could inhibit the proliferation of skin squamous cell carcinoma SCL-1 cells(the concentration of short peptides were 100 ng/mL).The 18-20 aa short peptides were divided into 3 segments,and their corresponding 6-7 aa short peptides were artificially synthesized.According to the above method,6-7 aa short peptides which could inhibit the proliferation of skin squamous cell carcinoma SCL-1 cells were screened out(the concentration of short peptides were 100 ng/mL).The 6-7 aa short peptides labeled with biotin were applied to local skin of nude mice.After 2 h,the local skin tissue was isolated and the skin permeability was observed by immunofluorescence.Results The 18-20 aa short peptide P1 and P2 could inhibit the proliferation of skin squamous cell carcinoma SCL-1 cells(P<0.05),and P3-P6 had no significant effect on proliferation of skin squamous cell carcinoma SCL-1 cell(P>0.05).The 6-7 aa short peptide P1-1,P1-2 and P2-2 corresponding to P1 and P2 could inhibit the proliferation of skin squamous cell carcinoma SCL-1 cells(P<0.05),and P1-3,P2-1,and P2-3 had no significant effect on the proliferation of skin squamous cell carcinoma SCL-1 cells(all P>0.05).Red fluorescence was detected in inside blood-vessel and vessel walls of nude mice skin applied with P1-1,P1-2 and P2-2 peptides.Conclusions The 6-7 aa PEDF short peptides P1-1,P1-2 and P2-2,which inhibit the proliferation of skin squamous cell carcinoma cells,are successfully screened out.They have good transdermal properties and are expected to be used in treatment of skin squamous cell carcinoma.

关 键 词：皮肤鳞癌 色素上皮衍生因子 短肽 透皮性 裸鼠 

分 类 号：R739.5[医药卫生—肿瘤]