Background::Non-coding RNAs have attracted considerable attention for their vital role in cancer. The purpose of this study was to determine the effects of non-coding RNAs on hepatocellular carcinoma (HCC) and reveal their regulatory mechanism in the pathophysiological process.Methods::We measured the expression of mucin 1 (MUC1) and miR-485-5p in tissues from 15 HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction and Western blot, screened for aberrantly expressed microRNAs (miRNAs) by miRNA microarrays. Bioinformatics tools were used to find the miRNA and circular RNA that regulated MUC1, which were validated by RNA immunoprecipitation assay and luciferase reporter assay. Cell counting kit-8, Transwell assays, and flow cytometry were used to conduct functional experiments. Proteins were examined by western blot and immunohistochemical staining assays. Significant differences between groups were estimated using the one-way analysis of variance. A P < 0.05 was considered statistically significant. Results::MUC1 was overexpressed in HCC tissues compared with that in paratumor tissues (normal vs. tumor, 1.007 ± 0.215 vs. 75.213 ± 18.403, t = 18.401, P < 0.001) while miR-485-5p was down-regulated (normal vs. tumor, 4.894 ± 0.684 vs. 1.586 ± 0.398, t= 16.191, P < 0.001). Inhibition of miR-485-5p promoted cell proliferation (73.33% ± 5.13% vs. 41.33% ± 3.51%, t= 8.913, P < 0.001), migration (102 ± 8 cells vs. 46 ± 8 cells, t= 8.681, P < 0.001), invasion (59 ± 7 cells vs. 28 ± 2 cells, t = 8.034, P < 0.01), and suppressed apoptosis (22.64% ± 6.97% vs. 36.33% ± 3.96%, t = 2.958, P < 0.05) of HepG2 cells with which MUC1 is knocked down. Mechanically, miR-485-5p binds to MUC1, while circHECTD1 binds to miR-485-5p, resulting in the indirect up-regulation of the MUC1 level. Conclusions::Our findings reveal that circHECTD1 facilitates HCC progression by sponging miR-485-5p to up-regulate MUC1.

目的:探讨犀地凉血方对HaCaT细胞LncRNA NEAT1/miR-485-5p/STAT3调控网络及细胞增殖、凋亡的影响。方法:以IL-17诱导的HaCaT细胞为研究对象，通过qPCR、Western印迹法检测LncRNA NEAT1、miR-485-5p、STAT3 mRNA和蛋白在HaCaT细胞和正常人表皮角质形成细胞（NHEK细胞）中的表达。采用荧光原位杂交技术（FISH）观察LncRNA NEAT1、miR-485-5p在HaCaT细胞中的表达；采用双萤光素酶报告基因实验验证LncRNA NEAT1、miR-485-5p、STAT3之间的靶向调控关系。犀地凉血方煎煮取汁给予大鼠灌胃后采集含药血清，以含药血清干预和/或LncRNA-NEAT1过表达载体转染HaCaT细胞，将HaCaT细胞分对照组、过表达LncRNA NEAT1组、犀地凉血方组、犀地凉血方+过表达LncRNA NEAT1组，采用qPCR、Western印迹法、流式细胞仪、CCK8等实验技术分别检测LncRNA NEAT1、miR-485-5p、STAT3表达及细胞增殖、凋亡情况。采用独立样本 t检验、单因素方差分析、LSD- t检验进行统计学分析。 结果:IL-17诱导的HaCaT细胞组LncRNA NEAT1、STAT3 mRNA相对表达水平（1.84 ± 0.21、2.20 ± 0.24）高于NHEK细胞组（1.00 ± 0.11、1.00 ± 0.11，均 P < 0.05），miR-485-5p相对表达水平（0.32 ± 0.04）低于NHEK细胞组（1.00 ± 0.12， t = 2.94， P = 0.015）；STAT3、p-STAT3蛋白表达水平（1.27 ± 0.13、2.43 ± 0.16）均高于NHEK细胞组（1.00 ± 0.11、1.00 ± 0.10， t = 2.54、3.02，均 P < 0.05）。FISH检测显示，miR-485-5p与LncRNA NEAT1共定位于HaCaT细胞质。双萤光素酶报告基因实验显示，共转染野生型LncRNA NEAT1、STAT3重组质粒时，miR-485-5p组细胞相对萤光素酶活性明显低于阴性对照组（均 P < 0.05）；而共转染突变型LncRNA NEAT1、STAT3重组载体质粒时，miR-485-5p组细胞萤光素酶活性与阴性对照组差异无统计学意义（均 P > 0.05）。过表达LncRNA NEAT1组HaCaT细胞中LncRNA NEAT1、STAT3（包括STAT3 mRNA和STAT3、p-STAT3蛋白）表达水平高于对照组，miR-485-5p表达水平低于对照组；犀地凉血方组LncRNA NEAT1、STAT3表达水平低于对照组，miR-485-5p表达水平高于对照组；犀地凉血方+过表达LncRNA NEAT1组LncRNA NEAT1、STAT3表达水平低于过表达LncRNA NEAT1组，而miR-485-5p表达水平高于过表达LncRNA NEAT1组（均 P < 0.05）。CCK8法检测显示，药物干预24、48、72 h时，过表达LncRNA NEAT1组HaCaT细胞增殖活性明显高于对照组，犀地凉血方+过表达LncRNA NEAT1组HaCaT细胞增殖活性高于犀地凉血方组，但二者均低于对照组（均 P < 0.05）。过表达LncRNA NEAT1组HaCaT细胞凋亡率（5.84% ± 0.28%）低于对照组（14.75% ± 0.83%，LSD- t = 3.48， P = 0.002），而犀地凉血方组（35.72% ± 3.62%）高于对照组（LSD- t = 5.34， P = 0.001）；犀地凉血方+过表达LncRNA NEAT1组细胞凋亡率（27.64% ± 2.82%）高于过表达LncRNA NEAT1组（LSD -t = 9.06， P < 0.001）。 结论:犀地凉血方能抑制HaCaT细胞增殖并促进其凋亡，作用机制与其干预LncRNA NEAT1/miR-485-5p/STAT3调控网络相关。

目的:探讨mTOR调节相关蛋白(regulatory-associated protein of mTOR,RAPTOR)对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)迁移、侵袭和增殖能力的影响.方法:利用TCGA生物信息数据库查询在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)组织与癌旁组织中差异表达的mRNA.West-ern blot检测RAPTOR在人口腔上皮细胞HOEC和OSCC细胞系中的表达.利用伤口愈合实验、Transwell实验、EdU实验检测各组细胞迁移、侵袭及增殖能力.生物信息学网站预测与RAPTOR靶向结合的微小RNA(mi-croRNA,miR).功能学实验验证miR-485-5p是否可以靶向RAPTOR影响OSCC细胞的迁移、侵袭和增殖.结果:RAPTOR在HNSCC组织中较癌旁组织表达增高.伤口愈合、Transwell和EdU实验结果示,RAPTOR有促进OSCC细胞的迁移、侵袭和增殖能力.miR-485-5p能与RAPTOR靶向结合,且miR-485-5p上调能逆转RAPTOR促进CAL27细胞迁移、侵袭和增殖能力.结论:miR-485-5p通过靶向RAPTOR抑制OSCC细胞迁移、侵袭和增殖能力.

目的:研究miR-485-5p在结直肠癌细胞中的表达水平,并探索其对结直肠癌细胞增殖和侵袭能力的影响.方法:运用荧光定量PCR法检测miR-485-5p在结直肠癌细胞株(HCT116和HCT8)及正常人类肠黏膜上皮细胞(FHC)中的表达水平.通过转染miR-485-5p mimic建立miR-485-5p过表达结直肠癌细胞,CCK-8实验及Transwell侵袭实验分别检测miR-485-5p对细胞增殖和侵袭能力的影响.生物信息学方法预测miR-485-5p下游靶基因并通过双荧光素酶报告基因检验、实时荧光定量PCR和Western blotting等方法进行实验验证.结果:miR-485-5p在结直肠癌细胞中的表达水平显著低于FHC细胞,差异有统计学意义(P＜0.05);miR-485-5p可抑制结直肠癌细胞在体外的增殖和侵袭能力;生物信息学结合双荧光素酶报告基因检测、实时荧光定量PCR及Western blotting等方法证实CD147为miR-485-5p调控的靶基因.结论:在结直肠癌中,miR-485-5p发挥了抑癌基因的功能,具有作为基因治疗的潜在价值.

目的:探讨长链非编码RNA LINC01001在乳腺癌组织中的表达及其对乳腺癌MCF-7细胞增殖能力的影响.方法:筛选2016年3月至2017年6月于湖北医药学院附属人民医院甲状腺乳腺外科行手术切除的乳腺癌患者癌及癌旁组织12例,qRT-PCR检测乳腺癌组织及癌旁组织LINC01001的相对表达量;通过重组质粒在人乳腺癌MCF-7细胞中过表达LINC01001,采用流式细胞术和MTT法检测过表达LINC01001后MCF-7细胞周期分布和增殖能力变化.qRT-PCR检测miR-485-5p和CDKN1AmRNA的表达变化,Western blotting检测CDKN1A、CDK4、CDK6和Cyclin D1蛋白表达变化.结果:LINC01001在乳腺癌组织中表达水平低于对应的癌旁组织(P＜0.01).LINC01001重组质粒转染MCF-7细胞后可显著抑制细胞周期进展(P＜0.05),抑制细胞的增殖能力(P＜0.05).过表达LINC01001后,MCF-7细胞的miR-485-5p的表达水平降低(P＜0.01),CDKN1A mRNA表达水平升高(P＜0.01);可促进CDKN1A蛋白的表达和抑制CDK4、CDK6和CyclinD1蛋白的表达.结论:LINC01 001在乳腺癌组织中表达降低,其可能通过下调miR-485-5p的表达上调CDKN1A的表达来抑制乳腺癌MCF-7细胞的增殖能力,为lncRNA的临床应用提供实验基础.