CAJ | 학술논문

背景与目的：中国是世界上食管癌的高发区，食管癌可发生早期淋巴结及血行转移，这使其表现出临床进展迅速并且预后很差的特性。研究显示癌组织侵袭和转移与其诱导产生的蛋白酶降解细胞外基质、基底膜能力密切相关，因此基质金属蛋白酶(matrix metalloproteinase，MMP)在肿瘤的发生、发展、转移过程中发挥重要作用，本文研究基质金属蛋白酶16(matrix metalloproteinase-16，MMP-16)在食管癌中的表达及其与食管癌浸润转移的关系，并为食管癌早期诊断及靶向治疗提供理论依据。方法：应用免疫组化、蛋白质印迹法(Western blot)及实时定量聚合酶链反应(real-time polymerase chain reaction，RT-PCR)检测食管癌及相应的正常组织的MMP-16蛋白表达水平和MMP-16 mRNA转录情况，并对其临床意义进行探讨。通过构建针对MMP-16的小片段发夹状RNA(shRNA)干扰表达质粒，应用划痕实验、Transwell小室实验、WST-1及流式细胞仪检测方法，研究shRNA表达质粒沉默MMP-16后对Eca109细胞的迁移、侵袭、增殖及凋亡水平的影响。采用t检验进行统计学分析。结果：在蛋白水平，MMP-16在癌组织和正常组织表达平均指数分别为0.569±0.380和1.483±0.982，差异有统计学意义(P<0.01)。MMP-16表达与食管鳞癌组织分化程度呈正相关(R=0.704，P<0.05)，与肿瘤的分期负相关(P<0.05)。干扰MMP-16的表达后Ec109细胞侵袭及迁移能力明显增加(P<0.05)，细胞凋亡水平下降，增殖差异无统计学意义(P>0.05)。结论：MMP-16在食管癌组织中低表达，随着细胞分化程度增加而表达增加，在Eca109细胞中干扰MMP-16后能够抑制肿瘤细胞的迁移、侵袭，并减少凋亡水平，MMP-16在食管癌中可能起保护作用。
배경여목적：중국시세계상식관암적고발구，식관암가발생조기림파결급혈행전이，저사기표현출림상진전신속병차예후흔차적특성。연구현시암조직침습화전이여기유도산생적단백매강해세포외기질、기저막능력밀절상관，인차기질금속단백매(matrix metalloproteinase，MMP)재종류적발생、발전、전이과정중발휘중요작용，본문연구기질금속단백매16(matrix metalloproteinase-16，MMP-16)재식관암중적표체급기여식관암침윤전이적관계，병위식관암조기진단급파향치료제공이론의거。방법：응용면역조화、단백질인적법(Western blot)급실시정량취합매련반응(real-time polymerase chain reaction，RT-PCR)검측식관암급상응적정상조직적MMP-16단백표체수평화MMP-16 mRNA전록정황，병대기림상의의진행탐토。통과구건침대MMP-16적소편단발협상RNA(shRNA)간우표체질립，응용화흔실험、Transwell소실실험、WST-1급류식세포의검측방법，연구shRNA표체질립침묵MMP-16후대Eca109세포적천이、침습、증식급조망수평적영향。채용t검험진행통계학분석。결과：재단백수평，MMP-16재암조직화정상조직표체평균지수분별위0.569±0.380화1.483±0.982，차이유통계학의의(P<0.01)。MMP-16표체여식관린암조직분화정도정정상관(R=0.704，P<0.05)，여종류적분기부상관(P<0.05)。간우MMP-16적표체후Ec109세포침습급천이능력명현증가(P<0.05)，세포조망수평하강，증식차이무통계학의의(P>0.05)。결론：MMP-16재식관암조직중저표체，수착세포분화정도증가이표체증가，재Eca109세포중간우MMP-16후능구억제종류세포적천이、침습，병감소조망수평，MMP-16재식관암중가능기보호작용。
Background and purpose:Esophageal carcinoma is one of main malignancies with rapid course and a poor prognosis in China. The reasons of poor overall survival are the invasion and metastasis of the tumor. Matrix metalloproteinase (MMPs) play essential roles in promoting tumor invasion and metastasis. In this study, we aimed to investigate the expression and functional signiifcance of matrix metalloproteinase 16(MMP-16) in esophageal squamous cell carcinoma (ESCC). We expect to ifnd a lead molecule for the beneift of early detecting tumor and the development of novel treatment of ESCC. Methods:The expression levels of MMP-16 protein and mRNA in human ESCC and the matched normal tissues were determined by immunohistochemistry, Western blot and Real-Time PCR (RT-PCR). The stable Ec109 cell line with MMP-16 knockdown and negative controls were established by RNA interference technology. The cell migration, invasion, proliferation and cell apoptosis of MMP-16 in stable interfered Ec109 cell line was examined by cell counting, scratch test, Transwell test and lfow cytometry assays. The data were analyzed by t test. Results:MMP-16 protein was downregulated in cancerous group compared with the matched normal tissue and correlated with the clinical features of histological differentiation (P<0.05) and tumor stage (P<0.05). The levels of MMP-16 mRNA and protein in Ec109 were signiifcantly decreased by RNA intetrence (P<0.05). We demonstrated that MMP-16 silencing signiifcantly promoted cell invasion and migration (P<0.05), and inhibited cell apoptosis (P<0.05), while no significant effect was observed on cell proliferation (P>0.05). Conclusion: MMP-16 is downregulated in human ESCC tissues. The cell migration and invasion is promoted by interference of MMP-16 in Ec109, while the cell apoptosis is inhibited. MMP-16 may be considered as a target gene for therapy of ESCC.