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刘 玮,宗佩君,许婷婷.LncRNA PCAT1 靶向 miR-329-3p 调控舌鳞状细胞癌细胞增殖、迁移侵袭和谷氨酰胺分解的机制研究[J].中国比较医学杂志,2021,31(11):102~107.
LncRNA PCAT1 靶向 miR-329-3p 调控舌鳞状细胞癌细胞增殖、迁移侵袭和谷氨酰胺分解的机制研究
Mechanism of lncRNA PCAT1 in regulating the proliferation, migration, invasion and glutamine decomposition of tongue squamous cell carcinoma cells by targeting miR-329-3p
投稿时间:2020-10-26  
DOI:10. 3969 / j.issn.1671-7856. 2021. 11. 015
中文关键词:  PCAT1  miR-329-3p  舌鳞状细胞癌  谷氨酰胺分解
英文关键词:PCAT1  miR-329-3p  tongue squamous cell carcinoma  glutamine decomposition
基金项目:
作者单位E-mail
刘 玮 潍坊护理职业学院病理教研室,山东 潍坊 262500 o1ggbo@ 163.com 
宗佩君 山东省潍坊市益都中心医院病理科,山东 潍坊 262500  
许婷婷 潍坊护理职业学院病理教研室,山东 潍坊 262500 258776507@ qq.com 
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中文摘要:
       目的 探讨长链非编码 RNA 前列腺癌相关转录物 1(PCAT1)调控舌鳞状细胞癌(TSCC)增殖、迁移侵袭和谷氨酰胺分解的调控机制。 方法 实时荧光定量聚合酶链反应( qRT-PCR)检测 TSCC 癌组织、癌旁组织、 TSCC 细胞和常口腔角质细胞中 PCAT1、miR-329-3p 的表达情况。 脂质体转染法将 pcDNA 组( 转染 pcDNA)、 pcDNA-PCAT1 组( 转染 pcDNA-PCAT1)、miR-NC 组( 转染 miR-NC)、miR-329-3p 组( 转染 miR-329-3p mimics)、 pcDNA-PCAT1+miR-NC 组( 共转染 pcDNA-PCAT1 和 miR-NC)、 pcDNA-PCAT1 + miR-329-3p 组( 共转染 pcDNA- PCAT1 和 miR-329-3p mimics)转染至 UM1 细胞;细胞计数试剂盒(CCK-8)、迁移实验(Transwell)、酶联免疫吸附试验(ELISA)检测细胞增殖、迁移侵袭和谷氨酰胺分解产物谷氨酸盐、α-酮戊二酸(α-KG)水平。双荧光素酶试验检测细胞中 PCAT1 与 miR-329-3p 之间的结合力。 结果 与癌旁组织组相比,癌组织组 PCAT1 表达明显上调,miR- 329-3p 表达明显下调;与 NHOK 组相比,UM1、CAL-27、SCC25 细胞中 PCAT1 表达均上调,miR-329-3p 表达均下调 (P<0. 05);与 pcDNA 组相比,pcDNA-PCAT1 组 PCAT1 表达明显上升,细胞增殖率、细胞侵袭数和细胞侵袭数均发生上调,谷氨酸盐水平和 α-KG 水平也发生明显升高(P<0. 05);PCAT1 可靶向调控 miR-329-3p;与 miR-NC 组相 比,miR-329-3p 组 TSCC 细胞 miR-329-3p 表达显著升高,细胞增殖率、细胞迁移数和细胞侵袭数均显著降低,谷氨酸盐水平和 α-KG 水平均明显降低(P< 0.05);与 pcDNA-PCAT1+miR-NC 组相比,pcDNA-PCAT1+miR-329-3p 组 PCAT1 表达显著降低,细胞增殖率、迁移细胞数和侵袭细胞数均明显降低,谷氨酸盐水平和 α-KG 水平也显著下调 (P<0. 05)。 结论 PCAT1 促进 TSCC 细胞增殖、迁移侵袭和谷氨酰胺分解,其机制与靶向 miR-329-3p 有关。
英文摘要:
       Objective To investigate the regulatory mechanism of long-chain non-coding RNA prostate cancer- related transcription 1 (PCAT1) in proliferation, migration, invasion and glutamine decomposition of tongue squamous cell carcinoma (TSCC) cells. Methods Real-time quantitative polymerase chain reaction was used to detect expression of PCAT1 and miR-329-3p in TSCC tissues, adjacent tissues, TSCC cells and normal oral keratinocytes. Liposomal transfection was applied to UM1 cells. Groups were the pcDNA group ( transfected with pcDNA), pcDNA-PCAT1 group (transfected with pcDNA-PCAT1), miR-NC group (transfected with miR-NC), miR-329-3p group (transfected with miR- 329-3p mimics), pcDNA-PCAT1 + miR-NC group (cotransfected with pcDNA-PCAT1 and miR-NC) and pcDNA-PCAT1 + miR-329-3p group ( cotransfected with pcDNA-PCAT1 and miR-329-3p mimics). Cell counting kit, migration assay (Transwell) and enzyme-linked immunosorbent assay were used to assess cell proliferation, migration / invasion, and levels of glutamine decomposition products glutamate and α-ketoglutarate (α-KG). The dual luciferase assay was used to detect binding between PCAT1 and miR-329-3p in cells. Results Compared with adjacent tissue, expression of PCAT1 in cancer tissue was upregulated significantly and expression of miR-329-3p was downregulated significantly. Compared with NHOK cells, PCAT1 expression was upregulated and miR-329-3p expression was downregulated in UM1, CAL-27 and SCC25 cells and of (P< 0. 05). Compared with the pcDNA group, expression of PCAT1 was significantly increased in the pcDNA- PCAT1 group, cell proliferation, invasion and migration were upregulated, and the levels of glutamate and α-KG were increased significantly (P<0. 05). PCAT1 targeted miR-329-3p. Compared with the miR-NC group, expression of miR- 329-3p in TSCC cells of the miR-329-3p group was increased significantly cell proliferation, migration, and invasion were reduced significantly, and the levels of glutamate and α-KG were reduced significantly ( P< 0. 05). Compared with the pcDNA-PCAT1+miR-NC group, expression of PCAT1 in the pcDNA-PCAT1+miR-329-3p group was reduced significantly, cell proliferation, migration, and invasion were reduced significantly, and the levels of glutamate and α-KG were also reduced significantly (P< 0. 05). Conclusions PCAT1 promotes the proliferation, migration, invasion, and glutamine decomposition of TSCC cells and its mechanism is related to targeting miR-329-3p.
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