卵巢癌细胞与卵巢癌淋巴定向高转移细胞分泌蛋白差异初筛  

Screening for lymphatic metastasis-associated differentially expressed secretory protein in ovarian carcinoma cell lines with directional highly lymphatic metastasis

作　　者：刘丽[1] 黎丹戎[1] 钟艳平[1] 李力[1] 高婷[1] 阮和云[1] 冠潇[1] 

机构地区：[1]广西医科大学附属肿瘤医院妇瘤科,广西南宁530021

出　　处：《中华肿瘤防治杂志》2013年第14期1053-1056,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然基金(81060218);教育部高等学校博士学科点专项科研基金(20050598002);广西自然科学基金(2012GX-NSFAA053157)

摘　　要：目的:分析卵巢癌细胞株(SKOV3)与卵巢癌淋巴结定向高转移细胞株(SKOV3-PM4)分泌蛋白的差异,寻找与卵巢癌淋巴定向转移密切相关的潜在诊断分子标志。方法:分别收集SKOV3与SKOV3-PM4培养上清,利用双向凝胶电泳(2-DE)及基质辅助激光解吸电离飞行时间质谱(MALDI_T0F-MS/MS)技术筛选及鉴定差异表达的蛋白点,对其进行生物学分析后采用蛋白质印迹法验证挑选目标蛋白在细胞分泌蛋白中的表达情况。结果:二维凝胶电泳显示,SK-OV3细胞分泌蛋白质点数约为569±156,SKOV3-PM4细胞分泌蛋白质点数约为598±175。选取差异较大的7个差异蛋白点进行MS+MS/MS质谱鉴定,分别为乙二醛酶1(GLO1)、翻译后调节肿瘤蛋白(TPTC)、二磷酸核苷激酶B(ND-PK-B)、膜联蛋白(ANXA1)、热休克蛋白27(HSP)、过氧化物氧化还原(PRDX1)和磷脂酰乙醇胺结合蛋白Ⅰ(PEBP-Ⅰ)。挑选NDPK-B、ANXA1和HSP蛋白经蛋白质印迹法验证发现,SKOV3-PM4细胞中ANXA1和HSP27表达高于SKOV3细胞,而NDPK-B表达低于SKOV3细胞。与质谱分析的结果基本一致。结论:在卵巢癌细胞与卵巢癌淋巴结定向高转移细胞之间分泌蛋白存在很大的差异,其中已被鉴定的ANXA1和HSP27与卵巢癌淋巴结转移密切相关,有望成为诊断或预测卵巢癌淋巴结转移患者血清检测的标志。OBJECTIVE：In order to screen for lymphatic metastasis-associated proteins and offer biomarkers and tar get gene to clinical diagnosis, the expression difference of secretory protein between ovarian carcinoma SKOV3 cells and o- varian carcinoma cell line with directional highly lymphatic metastasis SKOV3-PM4 cells were detected. METHODS：Cul- ture supernatant of SKOV3 and SKOV3-PM4 were collected respectively. Two-dimensional gel electrophoresis（2-DE） was used to isolate the total proteins. The differential protein spots were identified by matrix-assisted laser desorption/ioniza- tion time of flight mass spectrometry （MALDI-TOF-MS/MS）. The biological information of the proteins was analyzed. RESULTS： Secretory proteins of SKOV3 cells and skov3-pm4 cells were approximately 569 ＋ 156 and 598 ＋ 175, respec- tively. Seven different secretory proteins were selected and identified by MALDI-TOF-MS/MS. Those secretory proteins were Glyoxalase I translationally-controlled tumor protein, nucleoside diphosphate kinase B, annexin A1, heat shock pro- tein 27, peroxiredoxin 1, phosphtidylethanolamine-binding protein 1 respectively. The expression of nucleoside diphosphate kinase B,annexin Aland kteat shock protein 27 in culture supernatant were further validated by Western blot. The result is very similar to the result of MALDI-TOF-MS/MS. The express of ANXA1 and HSP27 in SKOV3-PM4 are higher than that of SKOV3,but the expression of NDPK-B in SKOVa-PM4 culture supernatant is lower than that of SKOV3. CON- CLUSIONS：There are many different secretory proteins between ovarian carcinoma SKOV3 cells and ovarian carcinoma cells. The secretory proteins of ANXA1 and HSP27 among identified proteins were related to ovarian carcinoma line with directional lymphatic metastasis and may become biomarkers for diagnoses and prognosticate of lymphatic metastasis in blood serum of ovarian carcinoma patient.

关 键 词：卵巢癌 淋巴定向转移 分泌蛋白 二维凝胶电泳 

分 类 号：R737.31[医药卫生—肿瘤]