CAJ | 학술논문

目的：探讨新发现的抑癌基因SRY盒包含基因30( SOX30)在结直肠癌中的表达及甲基化变化。方法：用逆转录PCR(RT-PCR)检测结直肠癌细胞株(HCT8、HCT116和SW480)中SOX30 mRNA的表达，利用去甲基化药物5-Aza-CdR处理结直肠癌细胞株72 h后检测SOX30基因高甲基化与其表达调控的关系。应用甲基化特异性PCR(MSP)检测上述3株结直肠癌细胞株及54例结直肠肿瘤组织和10例癌旁组织SOX30基因的甲基化发生情况，通过硫化测序PCR(BSP)验证结直肠癌组织和癌旁组织SOX30基因的甲基化改变，并对甲基化情况与结直肠癌患者病理特征进行相关性分析。结果：SOX30 mRNA在上述3株结直肠癌细胞株中表达与对照组相比均较低，且均发生明显高甲基化；去甲基化药物处理上述3株细胞株后， SOX30 mRNA的表达明显得到恢复。SOX30基因在大部分结直肠癌组织中均呈现高甲基化状态，甲基化发生率为79.6%(43/54)；而在相应的癌旁组织中甲基化检出率较低，仅为20%(2/10)。对结直肠癌患者中SOX30基因甲基化与其临床病理学资料进行相关性分析发现，SOX30基因高甲基化与患者的肿瘤病理分型显著相关(P=0.045)，而与患者的性别、年龄、吸烟与否、TNM分期和Ducks分期无显著相关性(P均>0.05)。结论：SOX30基因在结直肠癌中发生明显高甲基化修饰改变，其基因表达水平明显下调，而且其甲基化修饰率与患者肿瘤病理分型相关，提示SOX30可能在结直肠癌的发生过程中起重要作用。
목적：탐토신발현적억암기인SRY합포함기인30( SOX30)재결직장암중적표체급갑기화변화。방법：용역전록PCR(RT-PCR)검측결직장암세포주(HCT8、HCT116화SW480)중SOX30 mRNA적표체，이용거갑기화약물5-Aza-CdR처리결직장암세포주72 h후검측SOX30기인고갑기화여기표체조공적관계。응용갑기화특이성PCR(MSP)검측상술3주결직장암세포주급54례결직장종류조직화10례암방조직SOX30기인적갑기화발생정황，통과류화측서PCR(BSP)험증결직장암조직화암방조직SOX30기인적갑기화개변，병대갑기화정황여결직장암환자병리특정진행상관성분석。결과：SOX30 mRNA재상술3주결직장암세포주중표체여대조조상비균교저，차균발생명현고갑기화；거갑기화약물처리상술3주세포주후， SOX30 mRNA적표체명현득도회복。SOX30기인재대부분결직장암조직중균정현고갑기화상태，갑기화발생솔위79.6%(43/54)；이재상응적암방조직중갑기화검출솔교저，부위20%(2/10)。대결직장암환자중SOX30기인갑기화여기림상병이학자료진행상관성분석발현，SOX30기인고갑기화여환자적종류병리분형현저상관(P=0.045)，이여환자적성별、년령、흡연여부、TNM분기화Ducks분기무현저상관성(P균>0.05)。결론：SOX30기인재결직장암중발생명현고갑기화수식개변，기기인표체수평명현하조，이차기갑기화수식솔여환자종류병리분형상관，제시SOX30가능재결직장암적발생과정중기중요작용。
OBJECTIVE:To analyze the expression and methylation of SOX30 gene in colorectal cancer cell lines and colorectal cancer tissues.METHODS:ThemRNA expression and methylation ofSOX30 were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) andmethylation specific PCR (MSP). Demethylation experiment with 5-Aza-CdR was used to confirm the regulation ofSOX30 expression by DNA hypermethylation. The methylation ofSOX30 was further confirmed by sodium bisulfite DNA sequencing (BSP).RESULTS:Hypermethylation ofSOX30 gene was found in all the colorectal cancer cell lines,with reducedmRNAexpression. The mRNA expression ofSOX30 increased in colorectal cancer cell lines treated with 5-Aza-CdR,indicating thatSOX30 expression was regulated by DNA methylation.SOX30 hypermethylation was found in most of the colorectal cancer tissues (79.6%) ,but not in tumor adjacent tissues (20%). After investigating possible associations betweenSOX30 methylation and clinicopathologic features,we found thatSOX30 hypermethylation was associated with histological type,but not correlated to gender,age,smoking,clinical TNM stages and Ducks stages.CONCLUSION:Down-regulation ofSOX30 by hypermethylation was always found in colorectal cancer cell lines,and its hypermethylation was obviously associated with histological type,suggestingSOX30 playing an important role in the carcinogenesis of colorectal cancer.