CAJ | 학술논문

目的探讨跨膜丝氨酸蛋白酶4(transmembrane protease serine 4,TMPRSS4)在放疗后残余肝细胞癌转移中的作用及机制.方法建立具有肺转移潜能的人肝癌裸鼠原位模型,模拟临床分割放射治疗.将放疗后残余肝癌组织切除后再种植于正常裸鼠肝脏.6周后处死,测量放疗后再种植肝癌体积,计算其肝内移灶数目及肺转移率.HE染色观察放疗后肝癌组织学形态.Westernblot及RT-PCR检测放疗前后肝癌组织内上皮间质转化(epithelial-mesenchymal transition,EMT)相关基因及TMPRSS4的表达.结果对照组肝癌切除再种植后42 d,其肿瘤大小及肺转移率分别为(2.25士0.52)cm3和66.7％,而放疗后2d和放疗后30 d残癌切除再种植组,肿瘤大小分别为(1.61±0.51)cm3 (P＜0.05)和(2.60±0.61)cm3 (P＞0.05),肺转移率分别为12.5％ (P＜0.05)和100％ (P＜0.05).对照组及放疗后2d残癌切除再种植组无肝内转移灶,放疗后30 d残癌切除再种植组肝内转移灶数为18±8.05(P＜0.05).放疗后30 d残癌组织EMT相关蛋白N-cadherin、Vim-entin及TMPRSS4、SIP1的表达显著高于对照组及放疗结束后2d残癌组织,而E-cadherin表达则显著降低.结论放疗后残癌的转移潜能先降低后增强.放疗后残癌TMPRSS4表达增加进而诱导EMT的发生可能是后期转移潜能增强重要原因之一.
목적 탐토과막사안산단백매4(transmembrane protease serine 4,TMPRSS4)재방료후잔여간세포암전이중적작용급궤제.방법 건립구유폐전이잠능적인간암라서원위모형,모의림상분할방사치료.장방료후잔여간암조직절제후재충식우정상라서간장.6주후처사,측량방료후재충식간암체적,계산기간내이조수목급폐전이솔.HE염색관찰방료후간암조직학형태.Westernblot급RT-PCR검측방료전후간암조직내상피간질전화(epithelial-mesenchymal transition,EMT)상관기인급TMPRSS4적표체.결과 대조조간암절제재충식후42 d,기종류대소급폐전이솔분별위(2.25사0.52)cm3화66.7％,이방료후2d화방료후30 d잔암절제재충식조,종류대소분별위(1.61±0.51)cm3 (P＜0.05)화(2.60±0.61)cm3 (P＞0.05),폐전이솔분별위12.5％ (P＜0.05)화100％ (P＜0.05).대조조급방료후2d잔암절제재충식조무간내전이조,방료후30 d잔암절제재충식조간내전이조수위18±8.05(P＜0.05).방료후30 d잔암조직EMT상관단백N-cadherin、Vim-entin급TMPRSS4、SIP1적표체현저고우대조조급방료결속후2d잔암조직,이E-cadherin표체칙현저강저.결론 방료후잔암적전이잠능선강저후증강.방료후잔암TMPRSS4표체증가진이유도EMT적발생가능시후기전이잠능증강중요원인지일.
Objective To investigate the role and mechanism of TMPRSS4 in radiation induced metastasis of hepatocellular carcinoma (HCC).Methods Metastatic model of human HCC was established by orthotopic implantation of histologically intact human HCC tissue into the liver of nude mice.Mice bearing xenografts in liver were killed after radiation and the residual tumors were resected and reimplanted into the liver of normal nude mice.At the end of sixth week,the mice were killed and the histopathological features,tumor volume,intrahepatic and lung metastasis were evaluated.Expression of epithelial-mesenchymal transition (EMT) related genes including N-cadherin,Vimentin,SIP1 and TMPRSS4 were measured by Western blotting and RT-PCR.Results The tumor volume and frequency of lung metastasis of control group was 2.25±0.52 cm3 and 66.7％,respectively.Compared to control group,tumor diameter (1.61±0.51 cm3,P＜0.05) and lung metastasis (12.5％,P＜0.05) were significantly inhibited 2 days after radiation.Whereas,30 days after radiation,tumor growth recovered (2.60±0.61 cm3,P＞0.05) and lung metastasis was enhanced (100％,P＜0.05).There were no intrahepatic metastasis in the control group and in the group of reimplantation of HCC 2 days after radiation,while the tumors from those 30 days after radiation showed enhanced intrahepatic metastasis (18 ± 8.05,P＜ 0.01 ),with overexpression of SIP1,N-cadherin,Vimentin and TMPRSS4,and reduced expression of E-cadherin.Conclusion The metastasis potential of residual HCC after radiation was first inhibited and then promoted.Overexpression of TMPRSS4 plays a critical role in radiation induced long-term metastasis of HCC by facilitating EMT.