CAJ | 학술논문

目的观察微小RNA(miR)-483-5P在人乳腺癌组织和细胞株中表达,以及对人乳腺癌细胞增殖、细胞周期、凋亡和侵袭能力的影响.方法应用实时定量逆转录聚合酶链反应(RT-qPCR)检测乳腺癌组织、相应癌旁正常乳腺组织以及人乳腺癌细胞株MCF-7、MDA-MB-231和人乳腺浸润性导管癌旁皮肤细胞CCD-1095Sk中miR-483-5p表达；将miR-483-5p寡核苷酸通过脂质体转染MDA-MB-231细胞株,运用噻唑蓝(MTT)比色法观察其增殖,流式细胞仪分析细胞的周期、凋亡,Transwell实验观察其对细胞侵袭能力的影响.结果乳腺癌组织和癌旁正常乳腺组织中miR-483-5p的相对表达量分别为0.6333±0.1898和1.4471±0.3908,乳腺癌组织中miR-483-5p的表达明显低于正常组织(P＜0.05).乳腺癌细胞MCF-7、MDA-MB-231中miR-483-5p的表达也较正常乳腺细胞CCD-1095Sk显著降低,分别为其0.3290±0.0219和0.2307±0.0144倍.与对照组和无义序列转染组比较,miR-483-5p mimics转染组MDA-MB-231细胞增殖活性降低,增殖率为65.68％.Transwell实验结果显示,对照组及无义序列组穿膜细胞数分别为(203.8±12.0)个和(199.3±13.1)个,而转染miR-483-5p mimics组则为(89.8±12.4)个,较之对照组及无义序列组明显减少,差异有统计学意义(P＜0.05).转染miR-483-5p mimics后,细胞周期阻滞在G0/G1期为(55.1±5.2)％.结论 miR-483-5p在人乳腺癌组织和细胞株中低表达,体外实验初步证明miR-483-5p可抑制乳腺癌细胞株MDA-MB-231增殖和侵袭能力.
목적 관찰미소RNA(miR)-483-5P재인유선암조직화세포주중표체,이급대인유선암세포증식、세포주기、조망화침습능력적영향.방법 응용실시정량역전록취합매련반응(RT-qPCR)검측유선암조직、상응암방정상유선조직이급인유선암세포주MCF-7、MDA-MB-231화인유선침윤성도관암방피부세포CCD-1095Sk중miR-483-5p표체；장miR-483-5p과핵감산통과지질체전염MDA-MB-231세포주,운용새서람(MTT)비색법관찰기증식,류식세포의분석세포적주기、조망,Transwell실험관찰기대세포침습능력적영향.결과 유선암조직화암방정상유선조직중miR-483-5p적상대표체량분별위0.6333±0.1898화1.4471±0.3908,유선암조직중miR-483-5p적표체명현저우정상조직(P＜0.05).유선암세포MCF-7、MDA-MB-231중miR-483-5p적표체야교정상유선세포CCD-1095Sk현저강저,분별위기0.3290±0.0219화0.2307±0.0144배.여대조조화무의서렬전염조비교,miR-483-5p mimics전염조MDA-MB-231세포증식활성강저,증식솔위65.68％.Transwell실험결과현시,대조조급무의서렬조천막세포수분별위(203.8±12.0)개화(199.3±13.1)개,이전염miR-483-5p mimics조칙위(89.8±12.4)개,교지대조조급무의서렬조명현감소,차이유통계학의의(P＜0.05).전염miR-483-5p mimics후,세포주기조체재G0/G1기위(55.1±5.2)％.결론 miR-483-5p재인유선암조직화세포주중저표체,체외실험초보증명miR-483-5p가억제유선암세포주MDA-MB-231증식화침습능력.
Objective To explore the role of miR-483-5p in breast cancer,and investigate the expression of miR-483-5p in human normal breast skin cells (CCD-1095Sk),human breast cancer cell lines (MCF-7,MDA-MB-231),paired breast cancer tissues and adjacent non-tumor breast tissues.Methods The expression level of miR-483-5p in human normal breast skin cells (CCD-1095Sk),human breast cancer cell lines (MCF-7,MDA-MB-231),paired breast cancer tissues and adjacent non-tumor breast tissues was detected by using reverse transcription polymerase chain reaction (RT-PCR).MDA-MB-231 cells were transfected with miR-483-5p mimics and subjected to proliferation,cell cycle,apoptosis,and Transwell assays.Results The relative expression of miR-483-5p in human breast cancer specimens was 0.6333 ±0.1898,which was significantly lower than 1.4471 ±0.3908 of adjacent non-tumor breast tissues.miR-483-5p was also down-regulated in human breast cancer cell lines (MCF-7,MDA-MB-231) as compared with normal breast skin cells (CCD-1095Sk) (0.3290 ± 0.0219 and 0.2307 ± 0.0144 vs.1.0000 ±0.0000,both P ＜ 0.05).miR-483-5p mimics-transfected cells exhibited significantly reduced cell proliferation and the proliferation rate was 65.68％.Transwell assay revealed that the amount of invaded MDA-MB-231 cells in miR-483-5p transfected groups was significantly declined as compared with the control and scramble groups (89.8 ± 12.4 vs.203.8 ± 12.0 and 199.3 ± 13.1,both P ＜0.05).In comparison to control group,cell cycle was arrested in G0/G1 phase [(55.1 ± 5.2) ％] and there was no significant difference in apoptosis of cells transfected with miR-483-5p mimics.Conclusion miR-483-5p,which is down-regulated in human breast cancer specimens and cell lines,can suppress proliferation and invasive ability in vitro.