CAJ | 학술논문

背景:肝癌的转移复发被认为可能存在有肿瘤干细胞,后者在肿瘤细胞中数目极少,使得临床鉴别存在一定困难.Thy-1是卵圆细胞的表面标记蛋白,通过特异性标记物间接证明肿瘤中存在肿瘤干细胞是目前研究的重点.目的:探讨肝癌细胞中是否存在Thy-1标记阳性的卵圆细胞,并进行筛选鉴定.方法:流式细胞仪检测卵圆细胞标记蛋白Thy-1在肝癌细胞HepG2,BEL7402及正常肝细胞QSG7701的阳性表达情况,免疫磁珠分选Thy-1标记阳性的肝癌细胞,免疫荧光鉴定分选效率.裸鼠皮下接种实验设立3组,Thy-1~+细胞组裸鼠于背部皮下接种分选的Thy-1~+细胞5×10~5/只,HepG2细胞阳性对照组同法接种HepG2细胞0.5×10~7/只,Thy-1细胞阴性对照组同法接种分选的Thy-1-细胞0.5×10~7/只,1个月后观察各组成瘤性.结果与结论:卵圆细胞表面标记蛋白Thy-1在肝癌细胞HepG2,BEL7402中的表达明显高于正常肝细胞QSG7701(P＜0.05).Thy-1~+细胞组免疫荧光染色阳性率为85%,HepG2细胞阳性对照组为1%,Thy-1~-细胞阴性对照组几乎无免疫荧光表达.HepG2细胞阳性对照组、Thy-1~+细胞组5只裸鼠全部成瘤,且瘤体体积相对较大;而Thy-1~-细胞阴性对照组仅有1只裸鼠成瘤,瘤体体积明显小于前2组(F=144.568,P＜0.05),提示肝癌细胞中存在Thy-1标记阳性的卵圆细胞样干细胞,其可能是肝癌细胞中的肿瘤干细胞.
배경:간암적전이복발피인위가능존재유종류간세포,후자재종류세포중수목겁소,사득림상감별존재일정곤난.Thy-1시란원세포적표면표기단백,통과특이성표기물간접증명종류중존재종류간세포시목전연구적중점.목적:탐토간암세포중시부존재Thy-1표기양성적란원세포,병진행사선감정.방법:류식세포의검측란원세포표기단백Thy-1재간암세포HepG2,BEL7402급정상간세포QSG7701적양성표체정황,면역자주분선Thy-1표기양성적간암세포,면역형광감정분선효솔.라서피하접충실험설립3조,Thy-1~+세포조라서우배부피하접충분선적Thy-1~+세포5×10~5/지,HepG2세포양성대조조동법접충HepG2세포0.5×10~7/지,Thy-1세포음성대조조동법접충분선적Thy-1-세포0.5×10~7/지,1개월후관찰각조성류성.결과여결론:란원세포표면표기단백Thy-1재간암세포HepG2,BEL7402중적표체명현고우정상간세포QSG7701(P＜0.05).Thy-1~+세포조면역형광염색양성솔위85%,HepG2세포양성대조조위1%,Thy-1~-세포음성대조조궤호무면역형광표체.HepG2세포양성대조조、Thy-1~+세포조5지라서전부성류,차류체체적상대교대;이Thy-1~-세포음성대조조부유1지라서성류,류체체적명현소우전2조(F=144.568,P＜0.05),제시간암세포중존재Thy-1표기양성적란원세포양간세포,기가능시간암세포중적종류간세포.
BACKGROUND:There may be tumor stem cells during metastasis and recurrence of liver cancer.The number of tumor stem cells is very few,so it is difficult to be identified.Thy-1 is a surface marking protein of oval cells.Specific marker can indirectly confirm the existence of tumor stem cells in the tumor,which is a focus in recent studies.OBJECTIVE:To study whether Thy-1 positive marked oval cells was expressed in hepatoma carcinoma cells,and investigate its screening and identification.METHODS:Flow cytometry was used to identify that Thy-1 positive expression in hepatoma carcinoma cells HepG2 and BEL-7402 and normal hepatocytes QSG-7701.Immunomagnetic bead sorting was utilized to select hepatoma carcinoma cells with Thy-1.Immunofluorescence was used to identify selected efficiency.Naked mouse inoculated with hepatoma carcinoma cells in the skin test was assigned to three groups.In the Thy-1~+ cells group,naked mice were inoculated with Thy-1~+ cells (5×10~5/mouse)underneath the skin of the back.In the HepG2 cell positive control group,naked mice were inoculated with HepG2 cells(0.5×10~7/mouse)underneath the skin of the back.In the Thy-1' cells negative control group,naked mice were inoculated with Thy-1~- cells(0.5×10~7/mouse)underneath the skin of the back.Tumorigenicity in each group was observed one month later.RESULTS AND CONCLUSION:Thy-1 expression in hepatoma carcinoma cells HepG2 and BEL7402 was significantly greater compared with normal hepatocytes QSG7701(P＜ 0.05).Positive rate of immunofluorescence staining was 85% in the Thy-1~+ cell group,1% in the HepG2 cell positive control group,0% in the Thy-1~- cell negative control group.There were nubble of five mice in HepG2 cell positive control and Thy-1~+ groups,and the volume was large.However,there was one nubble in Thy-1~- group,and the volume was significantly small(F=144.568,P＜ 0.05).Results indicated that hepatoma carcinoma cells contained Thy-1 labeled positive oval cells-like stem cells,which may be tumor stem cells in hepatoma carcinoma cells.