摘要
目的研究组蛋白甲基化酶SETDB1在乳腺癌中的表达情况及对其癌细胞株增殖和迁移能力的影响。方法采用RT-PCR、Western blot和IHC方法检测38例乳腺癌与癌旁组织及癌细胞株中SETDB1的表达水平;采用稳定转染方法筛选出沉默SETDB1基因的克隆细胞株,采用CCK-8、软琼脂克隆形成及流式细胞术检测沉默SETDB1后对肿瘤细胞增殖能力的影响;采用Transwell方法检测沉默后对肿瘤细胞迁移能力的影响。结果 38例乳腺癌与癌旁组织中23例癌组织、13例癌旁组织阳性表达(P<0.05),SETDB1的mRNA水平相对表达量分别为(1.20±0.08)、(0.96±0.05)(t=2.44,P<0.05);沉默T47D中SETDB1基因后,GFP、sh2、sh4株形成克隆个数分别是(70.00±2.00)、(42.67±1.53)和(13.33±1.53)个(F=834.00,P<0.001);细胞周期中sh2株出现了G0/G1期阻滞,而sh4株出现了G2/M期阻滞(P<0.01);48 h穿膜个数分别是(54.25±1.71)、(16.50±1.29)、(9.00±0.82)个(F=1344.00,P<0.001)。结论 SETDB1在乳腺癌中高表达,沉默该基因可影响癌细胞株的增殖和迁移能力,有望作为乳腺癌诊断及治疗的新靶点。
Objective To study the histone methyhransferase SETDB1 expression in breast cancer and the proliferation and migration ability of breast cancer cells. Methods The SETDB1 expression on 38 cases of breast cancer tissues, cancer adjacent tissues and breast cancer cells were detected by RT-PCR, Western blot and IHC. The knockdown SETDB1 gene cell clones were screened out with stable transfection method, and the knockdown SET- DB1 tumor ceils proliferation ability was detected by CCK-8, clone forming and flow cytometry assay. The migration ability changes were detected by transwell assay. Results In 38 cases, 23 cases of carcinoma tissues and 13 cases of tissue adjacent to carcinoma were positively expressed （ P 〈 0. 05 ） , and the mRNA relative expression levels of SETDB1 were （1.20 ±0. 08）, （0. 96 ±0. 05） （t =2.44,P 〈0. 05）. The formed clone numbers in GFP, sh2, sh4 were （70.00 ±2. 00）, （42.67±1.53 ）, （ 13.33 ±1.53 ） （ F = 834.0, P 〈 0. 001 ）. sh2 blocked cell cycle in G0/G1 phase, while sh4 was arrested in G2/M phase. The numbers of migration respectively were （54.25 ± 1.71）, （16.50±1.29）, （9.00±0.82） （F=1344.00, P〈0.001）. Conclusion SETDB1 is frequently overexpressed in breast cancer. Knockdown SETDB1 can inhibit the proliferation and migration capacity of cancer cell, which indicates that SETDB1 is a new target for diagnosis and treatment of breast cancer.
出处
《安徽医科大学学报》
CAS
北大核心
2016年第8期1081-1087,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金联合项目(编号:U1204821)
国家自然科学基金面上项目(编号:81572794)
