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Abstract：
BACKGROUND: TMPO-AS1 is a recently characterized oncogenic lncRNA in ovarian cancer. Its role in other ovary diseases is unknown. This study explored its role in polycystic ovary syndrome (PCOS).
METHODS: Follicular fluid was extracted from both PCOS patients and controls. The levels of TMPO-AS1 and mature and premature miR-335-5p were analyzed by RT-qPCR. The role of TMPO-AS1 in regulating miR-355-5p maturation in granulosa-like tumor (KGN) cells was analyzed by overexpression experiments. The interaction between TMPO-AS1 and premature miR-335-5p was analyzed by RNA pull-down assay. The subcellular location of TMPO-AS1 in KGN cells was analyzed by nuclear fractionation assay. The role of TMPO-AS1 and miR-335-5p in KGN cell proliferation was analyzed by BrdU assay.
RESULTS: TMPO-AS1 was increased in PCOS, while mature miR-355-5p was decreased in PCOS. TMPO-AS1 overexpression decreased mature miR-355-5p level but increased premature miR-355-5p. TMPO-AS1 was localized in both nucleus and cytoplasm. TMPO-AS1 directly interacted with premature miR-355-5p in KGN cells. TMPO-AS1 increased KGN cell proliferation while miR-355-5p decreased cell proliferation. The co-transfection assay showed that TMPO-AS1 reduced the suppressive effects of miR-355-5p on cell proliferation.
CONCLUSIONS: TMPO-AS1 might suppress miR-335-5p maturation to participate in PCOS.
METHODS: Follicular fluid was extracted from both PCOS patients and controls. The levels of TMPO-AS1 and mature and premature miR-335-5p were analyzed by RT-qPCR. The role of TMPO-AS1 in regulating miR-355-5p maturation in granulosa-like tumor (KGN) cells was analyzed by overexpression experiments. The interaction between TMPO-AS1 and premature miR-335-5p was analyzed by RNA pull-down assay. The subcellular location of TMPO-AS1 in KGN cells was analyzed by nuclear fractionation assay. The role of TMPO-AS1 and miR-335-5p in KGN cell proliferation was analyzed by BrdU assay.
RESULTS: TMPO-AS1 was increased in PCOS, while mature miR-355-5p was decreased in PCOS. TMPO-AS1 overexpression decreased mature miR-355-5p level but increased premature miR-355-5p. TMPO-AS1 was localized in both nucleus and cytoplasm. TMPO-AS1 directly interacted with premature miR-355-5p in KGN cells. TMPO-AS1 increased KGN cell proliferation while miR-355-5p decreased cell proliferation. The co-transfection assay showed that TMPO-AS1 reduced the suppressive effects of miR-355-5p on cell proliferation.
CONCLUSIONS: TMPO-AS1 might suppress miR-335-5p maturation to participate in PCOS.
摘要：
背景：TMPO-AS1是最近在卵巢癌中表征的致癌lncRNA。其在其他卵巢疾病中的作用尚不清楚。本研究探讨了其在多囊卵巢综合征（PCOS）中的作用。
方法：从PCOS患者和对照组中提取卵泡液。通过RT-qPCR分析TMPO-AS1和成熟和过早的miR-335-5p的水平。通过过表达实验分析了TMPO-AS1在颗粒样肿瘤（KGN）细胞中调节miR-355-5p成熟中的作用。通过RNA下拉法分析TMPO-AS1与早熟miR-335-5p之间的相互作用。通过核级分试验分析TMPO-AS1在KGN细胞中的亚细胞定位。通过BrdU实验分析TMPO-AS1和miR-335-5p在KGN细胞增殖中的作用。
结果：TMPO-AS1在PCOS中增加，而成熟miR-355-5p在PCOS中降低。TMPO-AS1过表达降低了成熟miR-355-5p水平，但增加了过早的miR-355-5p。TMPO-AS1位于细胞核和细胞质中。TMPO-AS1在KGN细胞中直接与过早的miR-355-5p相互作用。TMPO-AS1增加KGN细胞增殖，而miR-355-5p降低细胞增殖。共转染实验显示TMPO-AS1降低了miR-355-5p对细胞增殖的抑制作用。
结论：TMPO-AS1可能抑制miR-335-5p成熟参与PCOS。
方法：从PCOS患者和对照组中提取卵泡液。通过RT-qPCR分析TMPO-AS1和成熟和过早的miR-335-5p的水平。通过过表达实验分析了TMPO-AS1在颗粒样肿瘤（KGN）细胞中调节miR-355-5p成熟中的作用。通过RNA下拉法分析TMPO-AS1与早熟miR-335-5p之间的相互作用。通过核级分试验分析TMPO-AS1在KGN细胞中的亚细胞定位。通过BrdU实验分析TMPO-AS1和miR-335-5p在KGN细胞增殖中的作用。
结果：TMPO-AS1在PCOS中增加，而成熟miR-355-5p在PCOS中降低。TMPO-AS1过表达降低了成熟miR-355-5p水平，但增加了过早的miR-355-5p。TMPO-AS1位于细胞核和细胞质中。TMPO-AS1在KGN细胞中直接与过早的miR-355-5p相互作用。TMPO-AS1增加KGN细胞增殖，而miR-355-5p降低细胞增殖。共转染实验显示TMPO-AS1降低了miR-355-5p对细胞增殖的抑制作用。
结论：TMPO-AS1可能抑制miR-335-5p成熟参与PCOS。
