lncRNA MEG3通过下调miR-7-5p表达对鼻咽癌细胞放射敏感性的影响

目的 探讨lncRNA MEG3对鼻咽癌细胞的放射敏感性影响及潜在作用机制。方法 实验设置过表达对照组、过表达MEG3组、抑制miR-NC组、抑制miR-7-5p组、过表达对照+4 Gy组、过表达MEG3+4 Gy组、抑制miR-NC+4 Gy组、抑制miR-7-5p+4 Gy组、过表达MEG3+过表达miR-NC组、过表达MEG3+过表达miR-7-5p组。采用qRT-PCR检测miR-7-5p和MEG3表达;细胞克隆形成实验检测鼻咽癌细胞放射敏感性;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测荧光活性。结果 MEG3在鼻咽癌组织和细胞系中低表达;过表达MEG3和抑制miR-7-5p表达可增加鼻咽癌细胞放射敏感性且促进细胞凋亡。MEG3可靶向调节miR-7-5p表达;过表达miR-7-5p逆转了过表达MEG3对鼻咽癌细胞放射增敏和促进细胞凋亡的作用。结论 过表达MEG3增加鼻咽癌细胞放射敏感性,并促进细胞凋亡,其机制可能与下调miR-7-5p有关。     

lncRNA MEG3通过调控miR-21表达增加肺癌细胞放射敏感性

目的 探讨长链非编码RNA (lncRNA) MEG3对肺癌细胞H1299的放射敏感性调控机制。方法 运用qRT-PCR法检测具有放射抗性的H1299细胞中MEG3、miR-21-5p的表达。将过表达对照组(转染pcDNA3.1)、过表达MEG3组(转染pcDNA3.1-MEG3)、抑制miR-NC组(转染anti-miR-NC)、抑制miR-21-5p组(转染anti-miR-21-5p)、过表达MEG3+过表达miR-NC组(转染pcDNA3.1-MEG3和miR-NC)、过表达MEG3+过表达miR-21-5p组(转染pcDNA3.1-MEG3和miR-21-5p mimics)均用脂质体法转染。克隆形成实验检测细胞存活分数,流式细胞术检测细胞凋亡,双荧光素酶报告基因检测实验检测细胞中MEG3与miR-21-5p的结合力。结果 与正常肺上皮细胞相比,H1299细胞中MEG3表达明显降低,miR-21-5p表达明显升高;过表达MEG3或抑制miR-21-5p均可促进H1299细胞凋亡,增强放射敏感性;MEG3可靶向调控miR-21-5p的表达。过表达miR-21-5p可逆转MEG3对H1299细胞放射增强作用。结论 lncRNA MEG3可增强H1299细胞放射敏感性,其机制也许可能与靶向miR-21-5p有关。     

LncRNA MEG3靶向miR-181a-5p调控宫颈癌细胞放射敏感性

目的 研究LncRNA MEG3对宫颈癌细胞放射敏感性的影响,并探讨其作用机制。方法 运用qRT-PCR法检测放射抗性和放射敏感性宫颈癌细胞中LncRNA MEG3的表达;将过表达对照组(转染pcDNA 3.1)、过表达LncRNA MEG3组(转染pcDNA 3.1-LncRNA MEG3)、抑制miR-NC组(转染anti-miR-NC)、抑制miR-181a-5p组(转染anti-miR-181a-5p)、过表达LncRNA MEG3+过表达miR-NC组(共转染pcDNA 3.1-LncRNA MEG3和anti-miR-NC)、过表达LncRNA MEG3+过表达miR-181a-5p组(共转染pcDNA 3.1-LncRNA MEG3和anti-miR-181a-5p),均用脂质体法转染至SiHa细胞;克隆形成实验检测细胞的存活分数;流式细胞术检测细胞的凋亡率;双荧光素酶报告基因检测实验检测细胞的荧光活性;Western blot检测细胞中PTEN、p-Akt、Akt的蛋白表达。结果 与放射敏感组相比,放射抗性宫颈癌组织中LncRNA MEG3的表达明显降低(P<0.05),其表达量与宫颈癌细胞的放射敏感性呈正相关;过表达LncRNA MEG3、抑制miR-181a-5p均可显著增强宫颈癌细胞SiHa放射敏感性,促进凋亡(P<0.05);野生型LncRNA MEG3细胞的荧光活性受miR-181a-5p的抑制。过表达miR-181a-5p逆转了LncRNA MEG3对宫颈癌细胞放射增敏和促凋亡作用及对PTEN/Akt信号通路的调控。结论 长链非编码RNA LncRNA MEG3可增强宫颈癌细胞放射敏感性,其机制可能与靶向miR-181a-5p调控PTEN/Akt 信号通路有关,可为提高宫颈癌的预后提供新方向。     

LncRNA UCA1对肺癌细胞放射敏感性影响及机制研究

目的 研究长链非编码RNA (LncRNA) UCA1对肺癌细胞增殖、凋亡及放射敏感性影响及其机制。方法 运用qRT-PCR法检测肺癌细胞A549、H1299和人正常肺细胞HBE中UCA1、miR-513a-5p表达。将si-con组(转染si-con)、si-UCA1组(转染si-UCA1)、miR-513a-5p组(转染miR-513a-5p mimics)、miR-NC组(转染miR-NC)、IR+si-con组(转染si-con+照射)、IR+si-UCA1组(转染miR-NC+照射)、IR+miR-513a-5p组(转染miR-513a-5p mimics+照射)、IR+miR-NC组(转染miR-NC+照射)、IR+si-UCA1+anti-miR-513a-5p组(共转染si-UCA1和anti-miR-513a-5p+照射)均用脂质体法转染至A549、H1299细胞,然后部分组进行4Gy照射。MTT法检测各组细胞增殖,克隆形成实验检测细胞增敏比,流式细胞术检测各组细胞凋亡,双荧光素没报告基因检测实验检测各组细胞的荧光活性。结果 与HBE细胞相比,A549、H1299细胞中UCA1表达显著升高(P<0.05),miR-513a-5p表达显著降低(P<0.05)。抑制UCA1、过表达miR-513a-5p均可明显抑制A549、H1299细胞增殖、促进凋亡、提高放射敏感性(放射增敏比为1.897、2.146和1.615、1.872)。miR-513a-5p可抑制野生型UCA1细胞的荧光活性,且UCA1可负向调控miR-513a-5p的表达。抑制miR-513a-5p可逆转抑制UCA1对细胞的放射敏感性的增强作用。结论 抑制LncRNA UCA1可增强放射对肺癌细胞敏感性,其机制可能与靶向抑制miR-513a-5p有关。     

lncRNA H19靶向miR-106a-5p调控乳腺癌细胞放射敏感性的分子机制

目的:研究lncRNA H19对乳腺癌细胞MCF-7增殖、凋亡及放射敏感性的影响,并探讨其机制。方法:运用qRT-PCR检测MCF-7细胞中H19、miR-106a-5p的表达;将si-NC组（转染si-con）、si-H19组（转染si-H19）、si-H19+anti-miR-NC组（si-H19和anti-miR-NC共转染）、si-H19+anti-miR-106a-5p组（si-H19和anti-miR-106a-5p共转染）,均用脂质体转染至MCF-7细胞,再用4 Gy放射照射;MTT法检测各组细胞的增殖情况;流式细胞术检测各组细胞的凋亡情况;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果:与对照组相比,放射照射组（IR）MCF-7细胞中H19显著降低,miR-106a-5p显著升高（P＜0.05）;敲减H19、过表达miR-106a-5p均可抑制MCF-7细胞增殖并促进凋亡,增强细胞的放射敏感性;H19靶向miR-106a-5p。抑制miR-106a-5p可逆转敲减H19对MCF-7细胞放射敏感性的增强作用。结论:敲减H19可抑制乳腺癌细胞增殖,促进凋亡,增强放射治疗的敏感性,其机制可能与靶向miR-106a-5p有关,可为提高乳腺癌的放射敏感性提供新靶点。     

miR-216a-5p靶向KLF12对肝癌细胞放射敏感性的影响

目的探究miR-216a-5p靶向Krüppel样转录因子12(KLF12)对肝癌细胞放射敏感性的影响。方法实时反转录PCR(RT-qPCR)检测人正常肝细胞L-02, 人肝癌细胞Huh7、HepG2、Hep3B、MHCC-97H中miR-216a-5p、KLF12 mRNA水平并比较。采用0、2、4、6、8 Gy X射线照射HepG2细胞2 h, 比较不同剂量照射组间的miR-216a-5p、KLF12 mRNA水平。采用质粒转染构建miR-216a-5p和/或KLF12过表达的HepG2细胞, 并给予4 Gy X射线照射2 h。相应的分组为miR-NC(对照)组、miR-216a-5p组、miR-216a-5p+NC组、miR-216a-5p+KLF12组、IR+miR-NC组、IR+miR-216a-5p组、IR+miR-216a-5p+NC组、IR+miR-216a-5p+KLF12组, 比较分组间的miR-216a-5p、KLF12 mRNA水平。克隆形成实验检测细胞放射敏感性;CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡。多组间比较采用单因素方差分析, 两两比较采用LS...     

过表达miR-486-5p增强鼻咽癌放射抗拒细胞株CNE-2R放射敏感性

目的 探讨鼻咽癌放射抗拒细胞株CNE-2R过表达miR-486-5p后的放射增敏效应。方法 采用microRNA(miRNA)测序和RT-qPCR检测miR-486-5p在鼻咽癌亲本细胞CNE2和放射抗拒细胞CNE-2R中的表达水平。使用miR-486-5p过表达及阴性对照慢病毒液感染CNE-2R细胞,分别定义为CNE-2R-Mimics组和CNE-2R-NC组,利用CCK-8实验、平板克隆形成实验检测细胞增殖能力和经不同照射剂量（0 Gy、2 Gy、4 Gy、6 Gy、8 Gy）X射线照射后的细胞活力,Annexin V-APC/7-AAD双染法流式细胞术检测细胞凋亡情况。结果 miRNA测序和RT-qPCR结果均显示CNE-2R细胞中的miR-486-5p表达水平低于亲本细胞CNE2。CCK-8实验结果显示,培养48 h、72 h、96 h后,过表达miR-486-5p的CNE-2R细胞增殖能力较CNE-2R-NC组减弱（均P<0.05）;经2 Gy、4 Gy、6 Gy、8 Gy照射剂量照射后,CNE-2R-Mimics组存活分数低于CNE-2R-NC组（均P<0.05...     

CircMATR3调节miR-129-5p/SOX2轴对鼻咽癌细胞CNE1恶性生物学行为的影响

目的:探讨CircMATR3调节miR-129-5p/SOX2轴对鼻咽癌细胞恶性生物学行为的影响。方法:以鼻咽癌细胞系CNE1为研究对象,将CNE1细胞分NC组、si-NC组、si-CircMATR3组、miR-NC组、miR-129-5p mimic组、si-CircMATR3+inhibitor NC组、si-CircMATR3+miR-129-5p inhibitor组、si-SOX2组。RT-qPCR检测细胞中CircMATR3、miR-129-5p、SOX2 mRNA的表达;Western blot检测蛋白表达;双荧光素酶报告实验验证CircMATR3与miR-129-5p、miR-129-5p与SOX2之间的靶向关系;克隆形成实验检测细胞克隆形成能力;Hoechst33342实验观察细胞形态学变化;Transwell小室实验检测细胞侵袭能力;流式细胞术检测细胞周期与细胞凋亡。结果:CircMATR3与miR-129-5p、miR-129-5p与SOX2之间存在靶向关系;沉默CircMATR3或过表达miR-129-5p或沉默SOX2可以抑制鼻咽癌细胞CNE1的克隆形成、侵袭...     

miR-485-3p通过调控ATR信号通路增强MGC803细胞放射敏感性研究

目的 研究miR-485-3p对胃癌细胞MGC803放射敏感性影响及其可能作用机制。方法 将miR-485-3p mimic或ATR siRNA转染MGC803细胞后X线照射,MTT、克隆形成实验和凋亡实验观察细胞生物效应(表达检测照射6 Gy,克隆形成实验0、2、4、6、8、10 Gy)。分别用RT-PCR和蛋白印迹法检测miR-485-3p和ATR表达水平,DIANA、TargetScan和miRanda软件预测和双荧光素酶实验验证miR-485-3p对ATR靶向作用。结果 MGC803细胞放射后miR-485-3p表达下调。miR-485-3p过表达降低了细胞增殖和存活分数,增加了细胞凋亡率。经靶基因预测软件发现ATR可能是miR-485-3p靶基因,双荧光素酶实验进一步验证了ATR是miR-485-3p直接靶点。miR-485-3p负调控ATR表达,转染ATR siRNA抑制ATR信号通路后放射敏感性升高。结论 miR-485-3p可能靶向ATR,通过抑制ATR信号通路活性调节MGC803细胞放射敏感性。     

lncRNA PTPRG-AS1通过靶向miR-124-3p调控肝癌细胞凋亡和放射敏感性的研究机制

目的:探讨lncRNA PTPRG-AS1对肝癌细胞凋亡和放射敏感性的影响及作用机制。方法:培养正常肝细胞L02和肝癌细胞HepG2、SMMC-7721和BEL-7402,qRT-PCR检测细胞中PTPRG-AS1和miR-124-3p表达水平。转染si-PTPRG-AS1、miR-124-3p mimics至HepG2细胞,抑制HepG2细胞中PTPRG-AS1表达或过表达miR-124-3p,流式细胞术检测细胞凋亡,克隆形成实验检测细胞放射敏感性,Western Blot法检测Bcl-2和Bax蛋白表达。生物信息学软件预测PTPRG-AS1与miR-124-3p存在互补的核苷酸序列,双荧光素酶报告基因实验验证PTPRG-AS1与miR-124-3p之间的调控关系。结果:与正常肝细胞L02相比,肝癌细胞HepG2、SMMC-7721和BEL-7402中PTPRG-AS1表达显著升高（P＜0.05）,miR-124-3p表达显著降低（P＜0.05）。抑制PTPRG-AS1或过表达miR-124-3p均可促进肝癌HepG2细胞的凋亡,增强肝癌HepG2细胞的放射敏感性,抑制Bcl-2蛋白表达,促进Bax蛋白表达。PTPRG-AS1负调控miR-124-3p表达。抑制miR-124-3p表达可部分逆转抑制PTPRG-AS1表达对肝癌HepG2细胞凋亡的促进作用以及放射敏感性的增强作用。结论:抑制PTPRG-AS1表达可能通过上调miR-124-3p表达促进肝癌细胞的凋亡并提高其放射敏感性,是肝癌治疗的潜在作用靶点。     

lncRNA MEG3 increases the radiosensitivity of lung cancer cells by regulating miR-21

Objective To investigate the regulatory mechanism of long-chain non-coding RNA (lncRNA) MEG3 on the sensitivity of lung cancer cell line H1299 to irradiation. Methods The expression of MEG3 and miR-21-5p in lung cancer cell line H1299 was detected by qRT-PCR. Overexpression control group (transfected with pcDNA3.1), MEG3 overexpression group (transfected with pcDNA3.1-MEG3), miR-NC inhibition group (transfected anti-miR-NC), miR-21-5p inhibition group (transfected with anti-miR-21-5p), MEG3 overexpression+miR-NC overexpression group (co-transfected with pcDNA3.1-MEG3 and miR-NC), MEG3 overexpression+miR-21-5p overexpression group (co-transfected with pcDNA3.1-MEG3 and miR-21-5p mimics) were all transfected into H1299 cells by liposome method treated with 4Gy irradiation. Cell survival fraction was detected by colony formation assay. Cell apoptosis was detected by flow cytometry. The binding of MEG3 to miR-21-5p in cells was assessed by dual luciferase reporter assay. Results Compared with normal lung epithelial cells, the expression of MEG3 was significantly decreased, whereas the expression of miR-21-5p was significantly increased in the radioresistant lung cancer cells H1299. Overexpression of MEG3 or inhibition of miR-21-5p could promote the apoptosis and enhance the radiosensitivity of H1299 cells. MEG3 could targetedly regulate the expression of miR-21-5p. Overexpression of miR-21-5p could reverse the enhanced radiosensitivity of MEG3 to H1299 cells. Conclusion LncRNA MEG3 can enhance the sensitivity of lung cancer cells H1299 to irradiation. The mechanism may be related to targeting miR-21-5p.     

LncRAN MEG3 regulates the radiosensitivity of cervical cancer cells by targeting miR-181a-5p

Objective To evaluate the effect of long-chain non-coding RNA MEG3(LncRNA MEG3) on the radiosensitivity of cervical cancer cells, and to explore its underlying mechanism. Methods The expression of LncRNA MEG3 in cervical cancer cells was detected by qRT-PCR. In the overexpression control group (transfected with pcDNA 3.1), LncRNA MEG3 overexpression group (transfected with pcDNA 3.1-LncRNA MEG3), miR-NC inhibition group (transfected with anti-miR-NC), miR-181a-5p inhibition group (transfected with anti-miR-181a-5p), LncRNA MEG3+miR-NC overexpression group (co-transfected with pcDNA3.1-LncRNA MEG3 and anti-miR-NC), LncRNA MEG3+miR-181a-5p overexpression group (co-transfected with pcDNA 3.1-LncRNA MEG3 and anti-miR-181a-5p), all plasmids were transfected into SiHa cells by liposome method. The cell survival fraction was assessed by colony formation assay. The cell apoptosis rate was evaluated by flow cytometry. The cell fluorescence activity was assessed by dual luciferase reporter assay. The expression levels of PTEN, p-Akt and Akt proteins were detected by Western blot. Results Compared with the radiosensitive group, the expression of LncRNA MEG3 was significantly down-regulated in radiation-resistant cervical cancer tissues (P<0.05), and its expression level was positively correlated with the sensitivity of cervical cancer cells. Overexpression of LncRNA MEG3 or inhibition of miR-181a-5p could significantly enhance the irradiation sensitivity and promote the apoptosis of cervical cancer cell line SiHa (both P<0.05). The fluorescence activity of wild-type LncRNA MEG3 cells was inhibited by miR-181a-5p. Overexpression of miR-181a-5p reversed the irradiation sensitization and pro-apoptosis effect of LncRNA MEG3 and the regulation of the PTEN/Akt signaling pathway on cervical cancer cell. Conclusion LncRNA MEG3 can enhance the sensitivity of cervical cancer cells to radiation exposure, probably by targeting the miR-181a-5p and regulating the PTEN/Akt signaling pathway, which will provide a new direction for improving clinical prognosis of cervical cancer patients.     

沉默lncRNA HOTAIR上调miR-17-5p表达增加U87R细胞放射敏感性

目的:探讨lncRNA HOTAIR对胶质瘤U87R细胞和移植瘤放射敏感性影响及潜在作用机制。方法:将阴性对照质粒、沉默HOTAIR质粒、过表达miR-NC质粒、过表达miR-17-5p质粒分别转染到U87R细胞中,记为沉默对照组、沉默HOTAIR组、过表达miR-NC组、过表达miR-17-5p组;以上各组细胞分别用...     

LncRNA LINC00261下调miR-620表达调控鼻咽癌放射敏感性实验研究

目的:探讨LINC00261对鼻咽癌放射敏感性影响及其作用机制。方法:用qRT-PCR检测放射敏感、放射抵抗鼻咽癌组织中miR-620和LINC00261的相对表达水平。采用0、2、4、6、8 Gy 60Coγ射线照射鼻咽癌细胞系6-10B、HNE-3细胞后,qRT-PCR法检测miR-620和LINC0...     

miR-34s增强结直肠癌细胞对放射敏感性的作用研究

目的:探讨miR-34s对结直肠癌细胞放射敏感性的影响以及在电离辐射致DNA损伤中的作用。方法:利用实时荧光定量PCR（real-time quantitative fluorescence PCR,qRT-PCR）检测miR-34a/b/c-5p在结直肠癌细胞中的表达水平,以及电离辐射后miR-34a/b/c-5p表达水平变化趋势。基于克隆形成法和单击多靶模型建立细胞存活曲线,分析miR-34a/b/c-5p对结直肠癌细胞放射敏感性的影响。过表达miR-34a/b/c-5p并进行照射,利用免疫荧光法检测照射后γH2AX焦点形成情况,进而分析miR-34a/b/c-5p对电离辐射致DNA损伤的作用。结果:miR-34a/b/c-5p在HCT116细胞中的表达水平明显高于HT29细胞（P＜0.05）。HCT116细胞经4 Gy照射后24 h内,miR-34a/b/c-5p表达水平呈双峰变化趋势。与miR-NC组相比,过表达miR-34a/b/c-5p可显著增加结直肠癌细胞的放射敏感性,miR-34a/b/c-5p组细胞平均致死剂量（D0）和准阈值剂量（Dq）均明显降低,且辐射增敏比（SER）明显增加。过表达miR-34a/b/c-5p可显著增加电离辐射诱导的DNA双链断裂（double strand breaks,DSBs）水平,照射后1 h和8 h γH2AX焦点数明显高于miR-NC组（P＜0.05）。结论:miR-34s为放射响应miRNA分子,过表达miR-34s可增加电离辐射诱导的DNA损伤水平并增强结直肠癌细胞的放射敏感性。     

miR-409-3 p调控靶基因Beclin-1抑制小细胞肺癌细胞的自噬进而抑制其生长和转移

目的:探究miR-409-3p及其靶基因对小细胞肺癌细胞自噬、生长和转移的影响.方法:qRT-PCR检测miR-409-3p在HPAEpiC、SBC-5、NCI-H446、HOP-92细胞中的表达,SBC-5细胞中转染miR-409-3p mimic和miR-NC,CCK8法检测转染后细胞增殖,Transwell小室法...