长链非编码RNA PCGEM1对肝癌Huh7细胞增殖、侵袭及凋亡的影响
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摘要
目的探讨长链非编码RNA(LncRNA) PCGEM1对肝癌细胞增殖、侵袭及凋亡的影响。方法构建LncRNA PCGEM1慢病毒沉默表达载体,培养人肝癌细胞系Huh7细胞至对数生长期,分为空白对照组、阴性对照组和观察组,空白对照组细胞未进行转染,阴性对照组细胞应用空慢病毒载体进行转染,观察组细胞应用LncRNA PCGEM1慢病毒沉默表达载体进行转染。采用实时荧光定量聚合酶链反应检测各组肝癌细胞中LncRNA PCGEM1及Caspase-3、Caspase-9 mRNA表达,细胞计数试剂盒-8检测细胞的增殖能力,Transwell小室实验检测细胞的迁移能力,流式细胞术检测细胞的凋亡情况。结果 3组细胞中LncRNA PCGEM1及Caspase-3、Caspase-9 mRNA相对表达量比较差异有统计学意义(F=4.328、5.473、4.198,P<0.05);观察组细胞中LncRNA PCGEM1及Caspase-3、Caspase-9mRNA相对表达量均显著低于空白对照组和阴性对照组(P<0.05),空白对照组与阴性对照组细胞中LncRNA PCGEM1及Caspase-3、Caspase-9 mRNA相对表达量比较差异无统计学意义(P>0.05)。转染24、48、72、96 h时,3组细胞增殖能力比较差异有统计学意义(F=4.126、4.572、5.218、5.379,P<0.05);观察组细胞增殖能力低于空白对照组和阴性对照组(P<0.05),空白对照组与阴性对照组细胞增殖能力比较差异无统计学意义(P>0.05)。3组细胞迁移细胞数、细胞凋亡率比较差异有统计学意义(F=6.873、6.844,P<0.05),观察组迁移细胞数及细胞凋亡率显著低于空白对照组和阴性对照组(P<0.05);空白对照组与阴性对照组迁移细胞数、细胞凋亡率比较差异无统计学意义(P>0.05)。结论肝癌Huh7细胞中存在LncRNA PCGEM1表达,下调LncRNA PCGEM1的表达能抑制肝癌细胞增殖、侵袭及凋亡。
Objective To investigate the effects of long noncoding RNA(LncRNA) PCGEM1 on the proliferation,invasion and apoptosis of hepatoma carcinoma cells. Methods LncRNA PCGEM1 lentivirus silencing expression vector was constructed,and human hepatoma carcinoma Huh7 cells were cultured. The Huh7 cells which in logarithmin growth phase were divided into blank control group,negative control group and observation group. The Huh7 cells in the blank control group were not transfected,while the Huh7 cells in the negative control group were transfected with empty lentivirus vector,the Huh7 cells in the observation group were transfected with LncRNA PCGEM1 lentivirus silencing expression vector. The expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells was detected by real-time fluorescence quantitative polymerase chain reaction. The proliferation ability of Huh7 cells was detected by cell counting kit-8.The migration ability of Huh7 cells was detected by Transwell assay. The apoptosis of Huh7 cells was detected by flow cytometry. Results There were significant differences in the relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells among the three groups(F=4.328,5.473,4.198; P<0.05). The relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells in the observation group was significantly lower than that in the blank control group and the negative control group(P<0.05),but there was no significant difference in the relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells between the blank control group and the negative control group(P>0.05). There were significant differences in the proliferation ability of Huh7 cells among the three groups at 24,48,72 and 96 hours after transfection(F=4.126,4.572,5.218,5.379; P<0.05); the proliferation ability of Huh7 cells in the observation group was lower than that in the blank control group and the negative control group(P<0.05),but there was no significant difference in the proliferation ability of Huh7 cells between the blank control group and the negative control group(P>0.05). There were significant differences in the number of migrating cells and apoptotic rate among the three groups(F=6.873,6.844; P<0.05); the number of migrating cells and apoptotic rate in the observation group were significantly lower than those in the blank control group and the negative control group(P<0.05); but there was no significant difference in the number of migrating cells and apoptotic rate between the blank control group and the negative control group(P>0.05). Conclusion LncRNA PCGEM1 is expressed in hepatoma carcinoma Hub7 cells,and down-regulation of LncRNA PCGEM1 can inhibit the proliferation,invasion and apoptosis of hepatoma carcinoma cells.
Objective To investigate the effects of long noncoding RNA(LncRNA) PCGEM1 on the proliferation,invasion and apoptosis of hepatoma carcinoma cells. Methods LncRNA PCGEM1 lentivirus silencing expression vector was constructed,and human hepatoma carcinoma Huh7 cells were cultured. The Huh7 cells which in logarithmin growth phase were divided into blank control group,negative control group and observation group. The Huh7 cells in the blank control group were not transfected,while the Huh7 cells in the negative control group were transfected with empty lentivirus vector,the Huh7 cells in the observation group were transfected with LncRNA PCGEM1 lentivirus silencing expression vector. The expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells was detected by real-time fluorescence quantitative polymerase chain reaction. The proliferation ability of Huh7 cells was detected by cell counting kit-8.The migration ability of Huh7 cells was detected by Transwell assay. The apoptosis of Huh7 cells was detected by flow cytometry. Results There were significant differences in the relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells among the three groups(F=4.328,5.473,4.198; P<0.05). The relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells in the observation group was significantly lower than that in the blank control group and the negative control group(P<0.05),but there was no significant difference in the relative expression of LncRNA PCGEM1 and Caspase-3,Caspase-9 mRNA in Huh7 cells between the blank control group and the negative control group(P>0.05). There were significant differences in the proliferation ability of Huh7 cells among the three groups at 24,48,72 and 96 hours after transfection(F=4.126,4.572,5.218,5.379; P<0.05); the proliferation ability of Huh7 cells in the observation group was lower than that in the blank control group and the negative control group(P<0.05),but there was no significant difference in the proliferation ability of Huh7 cells between the blank control group and the negative control group(P>0.05). There were significant differences in the number of migrating cells and apoptotic rate among the three groups(F=6.873,6.844; P<0.05); the number of migrating cells and apoptotic rate in the observation group were significantly lower than those in the blank control group and the negative control group(P<0.05); but there was no significant difference in the number of migrating cells and apoptotic rate between the blank control group and the negative control group(P>0.05). Conclusion LncRNA PCGEM1 is expressed in hepatoma carcinoma Hub7 cells,and down-regulation of LncRNA PCGEM1 can inhibit the proliferation,invasion and apoptosis of hepatoma carcinoma cells.
引文
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[3]RUHANEN H,HARIDAS P A N,ESKELINEN E L,et al.Depletion of TM6SF2 disturbs membrane lipid composition and dynamics in Hu H7 hepatoma cells[J].Biochim Biophys Acta Mol Cell Biol Lipids,2017,1862(7):676-685.
[4]肖雪珍,袁东方,辛丹丹,等.Aglaroxin C对肝癌细胞体内外生长的抑制作用[J].新乡医学院学报,2018,35(9):761-765.770.
[5]BASTOS V,FERREIRA-DE-OLIVEIRA J M P,CARROLA J,et al.Coating independent cytotoxicity of citrate-and PEG-coated silver nanoparticles on a human hepatoma cell line[J].J Environ Sci,2017,51(1):191-201.
[6]朱竹先,余晨,邱忠民,等.LncRNA PCGEM1和雄激素受体在前列腺癌中的共定位表达及临床意义[J].中国癌症杂志,2016,26(4):320-325.
[7]向菁,康丽华,管怀进.LncRNA在眼科疾病发生发展中的作用机制研究进展[J].眼科新进展,2017,37(6):579-582.
[8]BORNER T,ARNOLD M,RUUD J,et al.Anorexia-cachexia syndrome in hepatoma tumour-bearing rats requires the area postrema but not vagal afferents and is paralleled by increased MIC-1/GDF15[J].J Cachexia Sarcopenia Muscle,2017,8(3):417-427.
[9]李伟伟,耿晓松,孙建伟,等.靶向沉默DEK对肝癌Hep G2细胞增殖及细胞周期的影响[J].新乡医学院学报,2018,35(3):173-176.
[10]XU J,CHEN Y,QIAN L,et al.A Novel RNA-binding protein involves ABA signaling by post-transcriptionally repressing ABI2[J].Front Plant Sci,2017,8(24):1-12.
[11]李福兴,何薇,乔晓红,等.亲嗜性病毒整合位点1负向调控微小RNA-9与急性髓系白血病发病机制研究[J].中华实用儿科临床杂志,2018,33(9):693-696.
[12]FIORENZANO A,PASCALE E,GAGLIARDIM,et al.An ultraconserved element containing lncRNA preserves transcriptional dynamics and maintains ESC self-renewal[J].Stem Cell Reports,2018,10(3):1102-1114.
[13]HIRSCH G E,PARISII M M,MARTINS L A,et al.γ-oryzanol reduces caveolin-1 and PCGEM1 expression,markers of aggressiveness in prostate cancer cell lines[J].Prostate,2015,75(8):783-797.
[14]LI H,AN J,WU M,et al.LncRNA HOTAIR promotes human liver cancer stem cell malignant growth through downregulation of SETD2[J].Oncotarget,2015,6(29):27847-27864.
[15]ANDREIA S,MARC B,GEORGE C,et al.The clinical relevance of long non-coding RNAs in cancer[J].Cancers,2015,7(4):2169-2182.
[16]WANG Y,HE L,DU Y,et al.The long noncoding RNA lnc TCF7promotes self-renewal of human liver cancer stem cells through activation of wnt signaling[J].Cell Stem Cell,2015,16(4):413-425.
[17]KANG Y,SONG J,KIM D,et al.PCGEM1 stimulates proli-feration of osteoarthritic synoviocytes by acting as a sponge for miR-770[J].J Orthop Res,2015,34(3):412-418.
[18]ZHANG S,LI Z,ZHANG L,et al.MEF2-activated long noncoding RNA PCGEM1 promotes cell proliferation in hormonerefractory prostate cancer through downregulation of miR-148a[J].Mol Med Rep,2018,18(1):202-208.
[19]CHEN S,WANG L L,SUN K X,et al.LncRNA PCGEM1 induces ovarian carcinoma tumorigenesis and progression through Rho Apathway[J].Cell Physiol Biochem,2018,47(4):1578-1588.