摘要
目的:探讨Dickkopf相关蛋白3(DKK3)基因高甲基化对直肠癌增殖、迁移及侵袭的影响及其临床价值。方法:SW480、LOVO、HT29及HCT116等直肠癌细胞购自中国科学院细胞库。选择2016年1月至2016年12月直肠癌患者(直肠癌组)及直肠良性疾病患者(对照组)为研究对象。设计合成寡核苷酸序列(MON、UMON和CON)并转染至LOVO直肠癌细胞。采用甲基特异性PCR检测直肠癌患者(肿瘤组织、癌旁组织)、对照组直肠组织及直肠癌细胞系中DKK3基因甲基化状态;分析DKK3基因甲基化与临床病理因素、无进展生存期(PFS)及总生存期的关系;Transwell小室检测MON组、UMON组及CON组LOVO直肠癌细胞迁移及侵袭。两组间均数比较用独立样本t检验;多组间均数比较用单因素方差分析(组间比较用LSD-t检验)。Kaplan-Meier生存分析并采用Log-rank比较。结果:直肠癌组患者肿瘤组织DKK3基因甲基化率显著高于癌旁组织及对照组直肠组织(甲基化率分别72.2%、11.1%比12.0%),差异有统计学意义(χ^(2)=88.756,P<0.05)。SW480、HT29及HCT116等直肠癌细胞系中DKK3基因呈甲基化状态,而在LOVO直肠癌细胞中呈未甲基化状态。低+中分化、N2淋巴结转移、肿瘤最大径≥3 cm、T3+T4及Ⅲ+Ⅳ期患者DKK3基因甲基化率显著高于高分化、N0+N1淋巴结转移、肿瘤最大径<3 cm、T1+T2及Ⅰ+Ⅱ期患者(甲基化率分别79.31%比59.38%、86.11%比62.96%、83.67%比58.54%、83.33%比59.52%、82.76%比53.13%),差异有统计学意义(χ^(2)=5.922、4.673、5.833、5.198、7.610,均P<0.05)。DKK3基因甲基化患者PFS及中位生存期显著低于DKK3基因未甲基化患者[PFS为(24.4±3.8)个月比(33.7±4.1)个月,中位生存期为(31.5±4.2)个月比(42.8±5.4)个月],差异有统计学意义(Log-rank=21.135、30.876,P<0.05)。MON组LOVO直肠癌细胞1~6 d细胞活力、细胞迁移数及侵袭数均显著高于UMON组及CON组LOVO直肠癌细胞[1 d的吸光度(A)值分别为0.21±0.03、0.14±0.03�
Objective To study the effect of dickkopf-related protein 3(DKK3)gene’s hypermethylation on the proliferation,migration and invasion of rectal cancer.Methods Rectal cancer cell lines such as SW480,LOVO,HT29 and HCT116 were purchased from the cell bank of the Chinese academy of sciences.Patients with rectal cancer(rectal cancer group)and patients with benign rectal diseases(control group)from Jan 2016 to Dec 2016 were selected as the research subjects.Synthetic oligonucleotide sequences(MON,UMON and CON)were designed and transfected into LOVO cells.Methyl-specific PCR was used to detect the methylation status of DKK3 gene in rectal cancer patients(tumor tissue,adjacent tissue),control group rectal tissue and rectal cancer cell lines;DKK3 gene methylation and clinicopathological factors,progression-free survival(PFS)were analyzed.The relationship between PFS and median survival time;Transwell chamber was used to detect the migration and invasion of LOVO rectal cancer cells in MON group,UMON group and CON group.The independent samples t test was used for the comparison of means between two groups;the one-way analysis of variance was used for the comparison of means among multiple groups(LSD-t test was used for the comparison between groups).Kaplan-Meier survival analysis with Log-rank comparison.Results The methylation rate of DKK3 gene in tumor tissue of patients with rectal cancer was significantly higher than that of adjacent tissue and rectal tissue of control group(methylation rate was 72.2%,11.1%vs.12.0%,respectively),and the difference was statistically significant(χ^(2)=88.756,P<0.05).The DKK3 gene was methylated in rectal cancer cell lines in SW480,HT29 and HCT116,while was unmethylated in LOVO cells.The methylation rate of DKK3 gene in patients with low+moderate differentiation,N2 lymph node metastasis,tumor maximum diameter≥3 cm,T3+T4 andⅢ+Ⅳstages was significantly higher than that of well differentiated,N0+N1 lymph node metastasis,tumor maximum diameter<3 cm,T1+For patients with T2 andⅠ+Ⅱsta
作者
杜芹
王芳
王贵娟
孔庆丽
李鹏
刘鹏
冯丽娜
杨东
Du Qin;Wang Fang;Wang Guijuan;Kong Qingli;Li Peng;Liu Peng;Feng Lina;Yang Dong(Department of Oncology,Affiliated Hospital of Jining Medical College,Jining 272000,China;Department of Surgery,Affiliated Hospital of Jining Medical College,Jining 272000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第12期2460-2463,共4页
Chinese Journal of Experimental Surgery
基金
山东省自然科学基金(ZR2015AR012)。
