中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
3期
231-235
,共5页
李秋文%肖文华%萨仁高娃%董伟伟%赵惠霞%段昕妤%朱建华%康欢荣%付艳%郝怡鑫%王如良%宋林萍%叶明
李鞦文%肖文華%薩仁高娃%董偉偉%趙惠霞%段昕妤%硃建華%康歡榮%付豔%郝怡鑫%王如良%宋林萍%葉明
리추문%초문화%살인고왜%동위위%조혜하%단흔여%주건화%강환영%부염%학이흠%왕여량%송림평%협명
癌,肝细胞%抗原,肿瘤%组蛋白去乙酰化酶抑制剂%exosome
癌,肝細胞%抗原,腫瘤%組蛋白去乙酰化酶抑製劑%exosome
암,간세포%항원,종류%조단백거을선화매억제제%exosome
[Key words] Carcinoma,hepatocellular%Antigens,neoplasm%Histone deacetylase inhibitors%exosome
目的 探讨组蛋白去乙酰化酶抑制剂MS-275修饰的肝癌细胞exosome所含肿瘤抗原和免疫分子的变化. 方法 采用蔗糖梯度离心联合超滤技术分离和纯化肝癌HepG2细胞在MS-275处理前后分泌的exosome,并用免疫电镜和Western blot杂交技术分别观察exosome表达热休克蛋白70、人类白细胞抗原(HLA)-I、HLA-II、NY-ESO-1和CD80等免疫分子的变化.MS-275处理前后样本均数的比较用配对t检验.结果 经MS-275处理后,肝癌HepG2细胞分泌exosome的数量和总蛋白含量均较处理前显著增加,分别为(303.0±3.2)×106个/ml比(236.0±2.8)×106个/ml和(1.37±0.18) mg比(0.78±0.11)mg,t值分别为27.29和4.85,P值均＜0.01.免疫电镜下,100个exosome所含免疫分子HSP70、HLA-Ⅰ、HLA-DR和CD80胶体金颗粒数均明显增加,分别为(79.8±5.2)个比(60.2±3.2)个、(65.6±6.3)个比(38.4±2.7)个、(26.9±2.7)个比(3.0±0.16)个和(42.7±5.2)个比(28.9±1.9)个,t值分别为5.56、6.87、15.3和4.32,P值＜0.01或＜0.05；NY-ESO-1所含胶体金颗粒数无明显变化[(40.2±4.6)个比(32.2±4.5)个,t=2.15,P＞0.05].Western blot结果也证实了上述免疫电镜的发现.结论 MS-275修饰肝癌细胞能显著改变exosome各种免疫分子的含量和种类,有利于增强exosome的免疫刺激作用,为体外大规模制备抗肝癌亚细胞瘤苗提供了新思路.
目的 探討組蛋白去乙酰化酶抑製劑MS-275脩飾的肝癌細胞exosome所含腫瘤抗原和免疫分子的變化. 方法 採用蔗糖梯度離心聯閤超濾技術分離和純化肝癌HepG2細胞在MS-275處理前後分泌的exosome,併用免疫電鏡和Western blot雜交技術分彆觀察exosome錶達熱休剋蛋白70、人類白細胞抗原(HLA)-I、HLA-II、NY-ESO-1和CD80等免疫分子的變化.MS-275處理前後樣本均數的比較用配對t檢驗.結果 經MS-275處理後,肝癌HepG2細胞分泌exosome的數量和總蛋白含量均較處理前顯著增加,分彆為(303.0±3.2)×106箇/ml比(236.0±2.8)×106箇/ml和(1.37±0.18) mg比(0.78±0.11)mg,t值分彆為27.29和4.85,P值均＜0.01.免疫電鏡下,100箇exosome所含免疫分子HSP70、HLA-Ⅰ、HLA-DR和CD80膠體金顆粒數均明顯增加,分彆為(79.8±5.2)箇比(60.2±3.2)箇、(65.6±6.3)箇比(38.4±2.7)箇、(26.9±2.7)箇比(3.0±0.16)箇和(42.7±5.2)箇比(28.9±1.9)箇,t值分彆為5.56、6.87、15.3和4.32,P值＜0.01或＜0.05；NY-ESO-1所含膠體金顆粒數無明顯變化[(40.2±4.6)箇比(32.2±4.5)箇,t=2.15,P＞0.05].Western blot結果也證實瞭上述免疫電鏡的髮現.結論 MS-275脩飾肝癌細胞能顯著改變exosome各種免疫分子的含量和種類,有利于增彊exosome的免疫刺激作用,為體外大規模製備抗肝癌亞細胞瘤苗提供瞭新思路.
목적 탐토조단백거을선화매억제제MS-275수식적간암세포exosome소함종류항원화면역분자적변화. 방법 채용자당제도리심연합초려기술분리화순화간암HepG2세포재MS-275처리전후분비적exosome,병용면역전경화Western blot잡교기술분별관찰exosome표체열휴극단백70、인류백세포항원(HLA)-I、HLA-II、NY-ESO-1화CD80등면역분자적변화.MS-275처리전후양본균수적비교용배대t검험.결과 경MS-275처리후,간암HepG2세포분비exosome적수량화총단백함량균교처리전현저증가,분별위(303.0±3.2)×106개/ml비(236.0±2.8)×106개/ml화(1.37±0.18) mg비(0.78±0.11)mg,t치분별위27.29화4.85,P치균＜0.01.면역전경하,100개exosome소함면역분자HSP70、HLA-Ⅰ、HLA-DR화CD80효체금과립수균명현증가,분별위(79.8±5.2)개비(60.2±3.2)개、(65.6±6.3)개비(38.4±2.7)개、(26.9±2.7)개비(3.0±0.16)개화(42.7±5.2)개비(28.9±1.9)개,t치분별위5.56、6.87、15.3화4.32,P치＜0.01혹＜0.05；NY-ESO-1소함효체금과립수무명현변화[(40.2±4.6)개비(32.2±4.5)개,t=2.15,P＞0.05].Western blot결과야증실료상술면역전경적발현.결론 MS-275수식간암세포능현저개변exosome각충면역분자적함량화충류,유리우증강exosome적면역자격작용,위체외대규모제비항간암아세포류묘제공료신사로.
Objective To investigate the effects of the histone deacetylase inhibitor,MS-275,on the immune molecule content and categories in hepatocarcinoma exosomes.Methods Exosomes were isolated from the human hepatocarcinoma cell lines,HepG2 and Hep3b,and purified by a combination technique of ultrafiltration centrifugation and sucrose gradient ultracentrifugation.The expressions of heat shock protein (HSP)70,human leukocyte antigen (HLA)-I,HLA-DR,cluster of differentiation (CD) 80 and NY-ESO-1 on exosomes were analyzed with immunoelectron microscopy and Western blotting before and after MS-275 treatment.Intergroup differences were statistically analyzed by the Student's paired t-test.Results MS-275 treatment of both HepG2 and Hep3b cell types significantly increased the numbers of exosomes,their total protein content,and expression of HSP70,HLA-I and CD80 (per 100 exosomes),as compared to non-treated cells (all,P ＜ 0.01).MS-275 was also found to induce de novo expression of HLA-DR,but had no significant effect on NY-ESO-1 expression (P ＞ 0.05).The findings from immunoelecton microscopy confirmed those from Western blotting.Conclusion The histone deacetylase inhibitor,MS-275,can significantly alter the immune molecule content and categories in exosomes of hepatocarcinoma cells.The differential expression profile may reflect an anti-cancer immune response and represent molecular targets for novel anti-hepatoma therapeutic or preventative strategies.
