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。     

miR-485-3p通过靶向TLR1/NF-κB信号通路调节胃癌细胞放射敏感性

目的 研究miR-485-3p是否通过靶向TLR1对胃癌细胞放射敏感性起作用。方法分别用qRT-PCR和蛋白印迹法检测miR-485-3p和TLR1的表达变化,DIANA、TargetScan和miRanda软件预测和双荧光素酶报告实验验证miR-485-3p对TLR1的靶向作用。将miR-485-3p mimic或TLR1 siRNA转染到胃癌MGC803细胞中,放射处理细胞,凋亡实验、克隆形成实验和MTT检测细胞放射敏感性的变化。双荧光素酶报告实验检测miR-485-3p上调和TLR1沉默对NF-κB活性的影响。蛋白免疫印迹实验探究miR-485-3p上调和TLR1沉默对NF-κB靶基因的影响。结果 放射处理后胃癌细胞miR-485-3p表达下调,TLR1表达上调。靶基因预测软件发现TLR1可能是miR-485-3p的靶基因,双荧光素酶报告实验进一步验证了TLR1是miR-485-3p的直接靶点。miR-485-3p负调控TLR1的表达。miR-485-3p的过表达提高了细胞凋亡率,降低了细胞克隆形成和细胞群体增殖能力,增强了细胞的放射敏感性,且TLR1的沉默也具有相同作用。miR-485-3p上调和TLR1沉默均降低了NF-κB的活性,下调了NF-κB多个靶基因的表达。结论 miR-485-3p可能通过靶向TLR1调控NF-κB信号通路增强胃癌细胞的放射敏感性。     

miR-324-5p通过靶向调控DAPK1降低胰腺癌细胞的放射敏感性

目的:探讨死亡相关蛋白激酶1（death-associated protein kinase 1,DAPK1）在胰腺癌（pancreatic cancer,PaC）细胞放射敏感性中的作用,验证miR-324-5p通过靶向调控DAPK1影响胰腺癌细胞放射敏感性的机制。方法:通过生物信息学预测靶向DAPK1的miRNAs,并利用双荧光素酶报告基因检测miR-324-5p对DAPK1的调控作用。在PANC-1和MIA PaCa-2细胞中过表达miR-324-5p和DAPK1或抑制miR-324-5p后,对各细胞株进行放射诱导,检测细胞增殖和凋亡情况,以及凋亡相关分子的表达情况。结果:GEO数据集结果显示,胰腺癌组织中miR-324-5p的表达水平高于正常组织。与正常胰腺导管上皮细胞系（HPDE6-C7）相比,胰腺癌细胞系(Capan-1、Bxpc-3、PANC-1和MIA PaCa-2)中miR-324-5p表达水平更高（P均<0.001）。双荧光素报告基因检测结果表明,miR-324-5p靶向DAPK1的3' UTR,并且可下调DAPK1的表达。细胞实验结果证实,过表达miR-324-5p通过靶向调控DAPK1降低放射诱导的细胞凋亡和DNA的损伤,进而降低了胰腺癌细胞的放射敏感性。结论:miR-324-5p通过负调控DAPK1降低胰腺癌细胞对放射的敏感性,从而影响DNA修复和细胞凋亡。miR-324-5p/DAPK1途径可能为胰腺癌的靶向治疗提供了潜在的治疗靶点。     

Wnt10a及Wnt/β-catenin信号通路促进宫颈癌细胞增殖及其机制研究

目的:探讨Wnt10a及其Wnt/β-catenin信号通路在宫颈癌中的作用及其机制。方法:qPCR检测Wnt10a在正常宫颈细胞和不同宫颈癌细胞系中的表达,选取Wnt10a高表达细胞进行后续实验。构建Wnt10a过表达质粒,将细胞分为对照组、Wnt10a过表达组、LGK-974(Wnt抑制剂)组和Wnt10a过表达+LGK-974组。qPCR和Western blot检测细胞中Wnt10a mRNA和蛋白的表达,CCK-8检测细胞增殖能力,Western blot检测细胞中凋亡相关蛋白Bax、Bcl-2、Caspase-3和Wnt/β-catenin信号通路相关蛋白Wnt1、β-catenin、Cyclin D1、c-Myc的表达。结果:Wnt10a在C-33 A、Ca Ski、HeLa、HeLa 229宫颈癌细胞中的表达均显著高于正常宫颈细胞(P<0.05),且在C-33 A细胞中的表达水平最高。与对照组相比,Wnt10a过表达组细胞增殖能力、细胞中Wnt10a mRNA及蛋白水平显著升高(P<0.01),Bcl-2、Wnt1、β-catenin、Cyclin D1、c...     

miR-377-5p通过抑制AKT1/GSK-3β通路增强食管癌细胞TE-1的放射敏感性

目的 探究miR-377-5p对食管癌细胞TE-1放射敏感性的影响及机制。方法 TE-1细胞转染miR-377-5pmimic和miR-377-5pmimic NC构建过表达miR-377-5p细胞。对转染后的TE-1细胞照射后采用平板集落形成实验检测细胞放射生物学参数(D0、Dq、SF2),Transwell小室法检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,CCK8法检测细胞活力,流式细胞术检测细胞凋亡和细胞周期,免疫印迹检测AKT1和GSK-3β磷酸化水平。结果 2、4、6、8Gy照射的集落形成率均显著下降(P<0.05),且在同一剂量下miR-377-5pmimic组细胞集落形成率均显著低于miR-377-5pmimic NC组(P<0.05)。相比于miR-377-5pmimic NC组,miR-377-5pmimic组D0、Dq、SF2均显著下降(P<0.05),放射增敏比为1.34(D0值比);0 Gy照射后细胞侵袭、迁移、增殖能力显著下降,细胞凋亡水平显著上升,细胞周期被阻滞于G1期,AKT1和GSK-3β磷酸化水平显著下降(P<0.05);4 Gy照射后细胞侵袭、迁移、增殖能力下降,细胞凋亡水平显著上升,G1期显著延长,AKT1和GSK-3β磷酸化水平亦显著下降(均P<0.001)。结论 miR-377-5p能够增加食管癌细胞TE-1的放射敏感性,其机制可能是抑制AKT1/GSK-3β信号通路。     

基于miR-152-3p/Wnt轴辣椒素对乳腺癌他莫昔芬耐药细胞株LCC9耐药的作用机制

目的 探讨辣椒素对乳腺癌他莫西芬耐药细胞株LCC9耐药性的影响及其具体机制。方法 不同浓度辣椒素处理LCC9细胞,CCK8法检测细胞存活率,qRT-PCR检测MCF-7和LCC9细胞中miR-152-3p和Wnt1 mRNA相对表达水平。将LCC9细胞分为对照组、inhibitor NC组(转染inhibitor NC)、inhibitor组(转染miR-152-3p inhibitor)、inhibitor+辣椒素组(转染miR-152-3p inhibitor 24 h后,200μmol/L辣椒素处理),CCK8法检测细胞存活率、qRT-PCR检测miR-152-3p相对表达水平、流式细胞仪检测细胞凋亡率、双荧光素酶报告基因法检测miR-152-3p与Wnt的靶向关系、蛋白印迹法检测β-连环蛋白(β-catenin)和T细胞因子4(TCF4)蛋白相对表达量。结果 细胞存活率随辣椒素浓度的升高而降低(P<0.05)。LCC9细胞中miR-152-3p表达较MCF-7细胞降低,Wnt1 mRNA水平较MCF-7细胞升高(P<0.05)。与inhibitor NC组比较,in...     

宫颈癌ATM的表达与放射敏感性的关系

目的:探讨宫颈癌组织中ATM(毛细血管扩张性共济失调症突变蛋白,ataxia telangiectasia mutated)表达与放射敏感性的关系。方法:选择13例对放射耐受的宫颈癌患者作为实验组,同时选择34例对放射敏感的宫颈癌患者作为对照。采用免疫组织化学染色方法,对两组患者放疗前癌组织进行ATM蛋白检测。结果:癌组织ATM蛋白的表达在敏感组和耐受组分别为50.0%(17/34)和84.6%(11/13),耐受组明显高于敏感组(P=0.046)。结论:宫颈癌组织中ATM蛋白的高表达可导致肿瘤对辐射的耐受,ATM的表达状态可能作为预测宫颈癌放射敏感性的一项指标。     

miR-339-5p通过抑制NUDT5增强肺癌A549细胞的放射敏感性

目的：探讨miR-339-5p通过调控Nudix结构水解酶5（Nudix hydrolase 5,NUDT5）的表达对肺癌A549细胞放射敏感 性的影响。方法：体外低浓度梯度递增结合大剂量间断冲击方法诱导产生耐X射线的肺癌A549细胞株（RA549）,qPCR检测 miR-339-5p在人正常肺上皮细胞（BEAS-2B）、肺癌细胞系（A549、 L78、 H1299、 H460和RA549细胞）中的表达水平。根据对 RA549细胞的处理,实验分为NC组、 5 Gy组（5 Gy X 射线处理 RA549 细胞）、 5 Gy+miR-339-5p mimic组、 5 Gy+si-NUDT5组和 5 Gy+si-NUDT5+miR-339-5p inbibitor组,CCK-8法、Annexin V-FITC/PI双染流式术和WB分别检测各组细胞增殖、凋亡以及 NUDT5、 γ-H2AX 和 H2AX 蛋白表达的变化,双荧光素酶报告基因系统验证 miR-339-5p 和 NUDT5 的靶向关系。结果： miR-339-5p在各肺癌细胞系中表达显著低于BEAS-2B细胞（均P<0.05）,其中RA549细胞表达水平最低。验证显示NUDT5是 miR-339-5p的靶基因。与 NC 组相比, 5 Gy 组 RA549 细胞增殖活力和 NUDT5 表达显著降低（均 P<0.01）,凋亡率显著升 高（P<0.01）;与5 Gy组相比, 5 Gy+miR-339-5p mimic组RA549细胞的增殖活力显著降低（P<0.05）、凋亡率[（12.97±1.48） % vs （5.21±0.62） %, P<0.01]和γ-H2AX的表达水平（P<0.05）显 著 升 高 , 5 Gy+si-NUDT5 组 RA549 细 胞 中 NUDT5 的表达水平 （P<0.01）和细胞增殖活力（P<0.01）均显著降低,凋亡率[（11.21±1.06） % vs（5.54±0.44） %, P<0.01]和γ-H2AX表达水平（P<0.01）均 显著升高, 而5 Gy+si-NUDT5+miR-339-5p inhibitor组细胞中上述指标均回复至5 Gy组水平。结论：过表达miR-339-5p通过靶 向下调NUDT5表达肺癌A549细胞的放射敏感性。     

miRNA-15a通过靶向Wnt3a调控Wnt/β-catenin信号通路抑制卵巢癌的发展和转移

目的 :研究在卵巢癌发展转移进程中miRNA-15a的作用, 并探讨其作用机制。 方法 :收集锦州医科大学附属第一医院2012年5月至2015年6月32例卵巢癌患者的手术切除标本, 采用RT-PCR检测miRNA-15a表达。将培养的卵巢癌SKOV-3细胞分为转染miRNA-15a模拟物的实验组和转染无义miRNA的对照组, 采用CCK8、Transwell及流式细胞术检测细胞增殖、侵袭和凋亡的变化, 采用Western blot法检测相关蛋白表达。通过生物信息学方法预测miRNA-15a靶基因, 采用双荧光素酶报告基因实验结合Western blot法验证miRNA-15a对靶基因的调控。检测Wnt3a与miRNA-15a模拟物共转染SKOV-3细胞后对细胞增殖和侵袭的影响。 结果 :与癌旁组织中的miRNA-15a相对表达量0.692±0.228相比, 显著低于卵巢癌组织中的0.911±0.209, 差异具有统计学意义(P < 0.05)。与对照组细胞增殖能力相比, miRNA-15a过表达显著抑制实验组细胞增殖能力, 差异具有统计学意义(P < 0.05)。与对照组189.667±14.742相比, miRNA-15a过表达显著抑制实验组的细胞侵袭能力96.667±13.614, 差异具有统计学意义(P < 0.05)。miRNA-15a过表达实验组的细胞凋亡率27.400%±0.854%显著高于对照组细胞凋亡率6.933%±0.379%, 差异具有统计学意义(P < 0.05)。miRNA-15a模拟物下调Wnt3a和β-catenin的表达, 抑制细胞周期蛋白(Cyclin D1)和血管内皮生长因子(vascu-lar endothelial growth factor, VEGF)的表达, 促进E-钙离子依赖的细胞黏附素(E-calcium-dependent adhesion, E-cad)的表达。双荧光素酶报告基因实验和Western blot结果表明miRNA-15a靶向抑制Wnt3a表达。上调Wnt3a能显著降低miRNA-15a对卵巢癌细胞增殖、侵袭和对Wnt/β-catenin信号通路的抑制作用。结论:miRNA-15a通过靶向Wnt3a调控Wnt/β-catenin信号通路的激活以抑制卵巢癌的发展和转移。      

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.     

miR-377-5p enhances radiosensitivity of esophageal cancer cell line TE-1 by inhibiting AKT1/GSK-3β signaling pathway

Objective To investigate the effect and mechanism of miR-377-5p on the radiosensitivity of esophageal cancer cell line TE-1. Methods Esophageal cancer cell line TE-1 was transfected with miR-377-5p mimic and miR-377-5p mimic NC to construct the TE-1 cells overexpressing miR-377-5p. After the transfected TE-1 cells were exposed to ionizing radiation, radiobiological parameters(D0,Dq,SF2) were detected by plate colony formation assay. Cell invasion ability was assessed by Transwell chamber assay, Cell migration ability was evaluated by cell scratch assay, Cell viability was assessed by CCK8 assay. Cell apoptosis and cell cycle were detected by flow cytometry, Western blot. The phosphorylation levels of AKT1 and GSK-3βwere measured by Western blot. Results At the doses of 2, 4, 6, and 8 Gy, the colony formation rate in each group was significantly decreased (all P<0.05), and the colony formation rate at the same irradiation dose in the miR-377-5p mimic group was significantly lower than that in the miR-377-5p mimic NC group (P<0.05). Compared with the miR-377-5pmimic NC group, the D0, Dq and SF2 at 2Gy were decreased significantly in the miR-377-5pmimic group (all P<0.05), and the radiosensitization ratio(D0 ratio) was 1.34. After 0 Gy ionizing radiation, the invasion, migration and proliferation abilities of TE-1 cells were significantly decreased, the level of cell apoptosis was significantly increased, the cell cycle was arrested in G1 phase, and the phosphorylation levels of AKT1 and GSK-3β were significantly decreased in the miR-377-5pmimic group (all P<0.05). Following 4 Gy irradiation, cell invasion, migration, proliferation abilities were decreased, the level of cell apoptosis was increased significantly, the G1 phase was significantly extended and the phosphorylation levels of AKT1 and GSK-3β were also decreased significantly in the miR-377-5pmimic group (all P<0.001). Conclusion miR-377-5p can increase the radiosensitivity of esophageal cancer cell line TE-1, which may be due to the inhibition of the AKT1/GSK-3β signaling pathway.     

The effect of Wnt/β-catenin signaling on PD-1/PDL-1 axis in HPV-related cervical cancer

Infection with high-risk human papillomavirus (HPV), including HPV-16 and HPV-18, is the main cause of malignancies, such as cervical cancer. Viral oncoproteins encoded by HPV are expressed in HPV-positive cancers and associated with the early cancer stages and the transformation of normal cells. The signaling pathways involved in the transformation of normal cells to cancerous form and the subsequently expressed programmed cell death-ligand 1 (PD-L1) on the surface of the transformed cells lead to a disruption in recognition of tumor cells by the immune cell system, including T lymphocytes and dendritic cells which lead to the development of cervical cancer malignancy. These cells also produce modest levels of cytokines during exhaustion, tumor-infiltrating T CD4+ cells with high levels of PD-1 and CD39 release considerable quantities of cytokines. The Wnt/β-catenin signaling pathway, which controls the expression of genes involved in the tumor cells’ markers, is demonstrated to be one of the most potent cancer stimulants. It leads to the evasion of the tumor cells from immune cell detection and ultimately avoids being recognized by dendritic cells or T-cells. PD-L1, as an inhibitory immune checkpoint, is essential for controlling immune system activity by inhibiting T-cells’ inflammatory function. In the present review, we looked into how Wnt/β-catenin affects the expression of PD-L1 and related genes like c-MYC in cancer cells and its role in the development of HPV-induced malignancy. We hypothesized that blocking these pathways could be a potential immunotherapy and cancer prevention method.     

RNAi介导PEG10基因沉默对肝癌HepG2细胞Wnt/β-catenin信号通路及增殖的影响

目的 探讨PEG10基因对Wnt/β-catenin信号通路的影响在人肝癌发病机制中的作用,为肝癌的基因治疗提供新思路.方法 应用脂质体转染技术将靶向PEG10的短发夹状RNA(short hairpin RNA,shRNA)转染HepG2细胞,利用半定量RT-PCR分别检测PEG10、β-catenin和Wnt1基因...     

Upregulation of DKK3 by miR-483-3p plays an important role in the chemoprevention of colorectal cancer mediated by black raspberry anthocyanins

It is reported that black raspberry (BRB) anthocyanins could act as a potential chemopreventive agent for colorectal cancer (CRC). However, the underlying mechanism by which BRB anthocyanins inhibits the carcinogenesis of CRC cells has not been elucidated. The abnormal expression of microRNAs (miRNAs) that target important tumor suppressor genes is usually associated with CRC development. In this study, we explored whether BRB anthocyanins could affect the expression of certain miRNAs in an azoxymethane (AOM)/dextran sulphate sodium (DSS)-induced CRC mouse model and human CRC cell lines. miRNA microarray analysis was used to determine the differences in miRNA expression between AOM/DSS-induced mice fed with a diet supplemented without or with BRB anthocyanins. The expression of one particular miRNA, miR-483-3p, was found to decrease dramatically in AOM/DSS-induced mice that were fed with a diet supplemented with BRB anthocyanins. Subsequent quantitative real-time polymerase chain reaction and Western blot analyses showed that the reduced expression of miR-483-3p was accompanied by an increased expression of Dickkopf 3 (DKK3), a potential target of miR-483-3p as predicted by bioinformatic analysis. The protein and messenger RNA levels of DKK3 were significantly upregulated when the miR-483-3p level was reduced by a miR-483-3p-specific inhibitor, suggesting that DKK3 might be the target gene of miR-483-3p. In addition, the downstream factors of the DKK3 signaling pathway, which included Wnt/β-catenin, also played a role in the miR-483-3p-mediated anticancer effect of BRB anthocyanins. Thus, miR-483-3p might be a potential target in BRB anthocyanin-mediated prevention of CRC.     

miR-1471对胶质瘤细胞增殖、侵袭和迁移能力的影响及其机制探讨

目的:探讨miR-1471对胶质瘤细胞增殖、侵袭和迁移能力的影响及其机制。方法:体外培养人胶质瘤细胞U251,将miR-1471模拟物或阴性对照分别转染至细胞中,记为miR-1471组和阴性对照（NC组）,同时设置对照组。qRT-PCR检测转染效果。CCK8检测各组细胞增殖能力。Transwell实验检测各组细胞侵袭能力。划痕实验检测各组细胞迁移能力。qRT-PCR和Western blot检测转移黏附基因（metadherin, MTDH）及Wnt/β-catenin信号通路相关基因和蛋白的表达。结果:与对照组和NC组比较,miR-1471组细胞增殖活性均显著降低（P＜0.05）,侵袭细胞数均显著减少（P＜0.05）,划痕间距明显较大,细胞中MTDH、Wnt1和β-catenin mRNA和蛋白表达水平均显著降低（P＜0.05）。结论:miR-1471能抑制人胶质瘤细胞U251的增殖、侵袭和迁移能力,其作用机制可能与下调MTDH表达,抑制Wnt/β-catenin信号通路的激活有关。     

Let-7b-5p inhibits colon cancer progression by prohibiting APC ubiquitination degradation and the Wnt pathway by targeting NKD1

Naked cuticle homolog 1 (NKD1), which is expressed at low levels in many tumors, is considered an inhibitor of the Wnt/β-catenin pathway, but it is highly expressed in colon cancer and can promote colon cancer cell proliferation. miRNAs are involved in the occurrence and progression of many tumors. However, miRNAs that can regulate NKD1 and the mechanisms by which NKD1 regulates tumor progression remain ambiguous. This research aims to reveal the potential regulatory network of NKD1 in colon cancer. miRNA data downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were analyzed by bioinformatics to screen for potential miRNAs targeting NKD1. Let-7b-5p was found to inhibit proliferation, migration, and invasion of colon cancer cells targeting NKD1. Further studies suggested that let-7b-5p can modulate Wnt signaling activity, and the nuclear accumulation of β-catenin was significantly restrained by let-7b-5p through targeting NKD1. Moreover, NKD1 could prohibit the expression of the APC protein. Further studies manifested that NKD1 bound to APC and promoted the ubiquitination degradation of APC through restraining the expression of the deubiquitinating enzyme USP15 and blocking the combination between USP15 and APC. Functionally, NKD1 enhanced the proliferation and migration of colon cancer cells by inhibiting APC expression. This research revealed a novel mechanism by which the let-7b-5p-NKD1-APC-β-catenin signaling pathway inhibited colon cancer cell progression.     

阻断CLC-3 氯通道对结直肠癌细胞株生存率和侵袭转移能力的影响及其分子机制*

目的:通过研究CLC-3 在结直肠组织中的表达,探讨CLC-3 对结直肠癌细胞株SW480、SW620 的细胞生存率、侵袭转移能力的影响及其潜在的机制。方法:采用RT-PCR 方法测定不同分期结直肠癌组织和正常结直肠组织中CLC-3 的mRNA 表达水平。运用CLC-3 阻断剂DIDS、NPPB阻断SW480、SW620 细胞的CLC-3 表达后,采用CCK-8 法实验检测细胞生存率,细胞侵袭实验检测细胞侵袭转移情况;并采用免疫印迹法测定阻断CLC-3 表达后SW480、SW620 细胞Wnt/ β -catenin 信号通路相关蛋白表达。结果:结直肠癌组织中CLC-3 mRNA 表达水平高于结直肠炎黏膜组织和正常结直肠组织（P < 0.05）,且CLC-3 mRNA 表达和结直肠分期相关。抑制CLC-3 表达后,SW480、SW620 细胞的生存率和侵袭转移能力降低（P < 0.05）,且β -catenin、C-myc 、cyclinD 1、Ki-67、survivin表达降低（P < 0.05）。 结论:CLC-3 高表达与结直肠的发生发展有潜在联系,其机制可能与Wnt/ β -catenin 有关,为CLC-3 作为治疗结直肠癌的潜在新靶点提供实验基础。