CAJ | 학술논문

背景与目的：miR-101在胃癌、结肠癌、乳腺癌以及前列腺癌中表达下调，有类似抑癌基因样作用，然而，其在卵巢癌中的作用尚未明确。该研究旨在探讨miR-101是否通过靶向调控甲基化转移酶3A(DNMT3A)抑制人卵巢癌细胞生长与侵袭，从而进一步揭示miR-101的抑瘤机制。方法：采用实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction，qRT-PCR)检测22例卵巢癌组织及癌旁正常卵巢组织中miR-101的表达改变；将miR-101 mimics转染于卵巢癌SKOV3细胞，以DNMT3A siRNA为阳性对照，采用蛋白[质]印迹法(Western blot)检测外源过表达miR-101对DNMT3A蛋白表达水平的影响；采用噻唑蓝(thiazolyl blue，MTT)和Transwell侵袭实验检测外源高表达miR-101对人卵巢癌细胞生长与侵袭能力的影响。结果：qRT-PCR检测结果显示，miR-101在22例卵巢癌组织中的表达水平较癌旁正常组织明显下调；Western blot检测结果显示，外源过表达miR-101或沉默DNMT3A能下调SKOV3细胞DNMT3A蛋白的表达水平；MTT检测结果显示，转染miR-101 mimics或沉默DNMT3A 48、72和96 h后D值与对照组比较明显减少，差异均有统计学意义(P<0.05)；Transwell侵袭实验显示，转染miR-101 mimics或沉默DNMT3A 36 h后穿过基底膜的细胞数分别为(105±7)个和(107±13)个，与对照组(213±11)个比较能明显减缓SKOV3细胞的穿膜能力，差异有统计学意义(P<0.05)。结论：miR-101通过靶向调控DNMT3A抑制人卵巢癌细胞生长与侵袭。
배경여목적：miR-101재위암、결장암、유선암이급전렬선암중표체하조，유유사억암기인양작용，연이，기재란소암중적작용상미명학。해연구지재탐토miR-101시부통과파향조공갑기화전이매3A(DNMT3A)억제인란소암세포생장여침습，종이진일보게시miR-101적억류궤제。방법：채용실시정량취합매련식반응(quantitative real-time polymerase chain reaction，qRT-PCR)검측22례란소암조직급암방정상란소조직중miR-101적표체개변；장miR-101 mimics전염우란소암SKOV3세포，이DNMT3A siRNA위양성대조，채용단백[질]인적법(Western blot)검측외원과표체miR-101대DNMT3A단백표체수평적영향；채용새서람(thiazolyl blue，MTT)화Transwell침습실험검측외원고표체miR-101대인란소암세포생장여침습능력적영향。결과：qRT-PCR검측결과현시，miR-101재22례란소암조직중적표체수평교암방정상조직명현하조；Western blot검측결과현시，외원과표체miR-101혹침묵DNMT3A능하조SKOV3세포DNMT3A단백적표체수평；MTT검측결과현시，전염miR-101 mimics혹침묵DNMT3A 48、72화96 h후D치여대조조비교명현감소，차이균유통계학의의(P<0.05)；Transwell침습실험현시，전염miR-101 mimics혹침묵DNMT3A 36 h후천과기저막적세포수분별위(105±7)개화(107±13)개，여대조조(213±11)개비교능명현감완SKOV3세포적천막능력，차이유통계학의의(P<0.05)。결론：miR-101통과파향조공DNMT3A억제인란소암세포생장여침습。
Background and purpose:miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods:Quantitative real-time palymerase chain reaction (qRT-PCR) method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expres-sion of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays.Results:qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that theD values were signiifcantly lower than the control group, (P<0.05). After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was (105±7) and (107±13), respectively, which are signiifcantly different from the control group (213±11), indicating invasion of SKOV3 cells signiifcantly slowed down (P<0.05).Conclusion:miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.