The roles of miR-200c in colon cancer and associated molecular mechanisms

The expression of miR-200c has been widely reported to be elevated in tumor tissues and sera of patients with colorectal cancer (CRC) and has been found to correlate with poor prognosis. However, how miR-200c regulates the apoptosis, survival, invasion, metastasis, and tumor growth in colon cancer cells remains to be fully elucidated. This study seeks to further investigate the role of miR-200c in colon cancer development. The expression of miR-200c in tumor and peritumoral tissues of 101 colon cancer patients was measured by real-time PCR. miR-200c expression in HCT-116 and HT-29 colon cancer cells was silenced by adenovirus-carried expression of antisense mRNA against miR-200c. The protein levels of PTEN, p53 Ser¹⁵, PP1, and activated caspase-3 in HCT-116 and HT-29 cells were measured by Western blot. This study demonstrated that the expression of miR-200c was significantly higher in tumor tissues than in peritumoral tissues of colon cancer patients. The elevated miR-200c expression significantly correlated with the TNM stage, lymph node metastasis, and invasion of colon cancer. Silencing miR-200c expression significantly induced cell apoptosis, inhibited long-term survival, invasion, and metastasis, and delayed xenograft tumor growth. Importantly, silencing miR-200c expression sensitized the therapeutic effect of Ara-C (Cytarabine). The effects of silencing miR-200c expression were associated with upregulation of PTEN protein and p53 Ser¹⁵ phosphorylation levels in HCT-116 cells and PTEN protein expression in HT-29 cells. In conclusion, miR-200c functions as an oncogene in colon cancer cells through regulating tumor cell apoptosis, survival, invasion, and metastasis as well as xenograft tumor growth through inhibition of PTEN expression and p53 phosphorylation.     

miR-188-5p promotes oxaliplatin resistance by targeting RASA1 in colon cancer cells

The efficacy of chemotherapy for colon cancer is limited due to the development of chemoresistance. MicroRNA (miR)-188-5p is downregulated in various types of cancer. The aim of the present study was to explore the molecular role of miR-188 in oxaliplatin (OXA) resistance. An OXA-resistant colon cancer cell line, SW480/OXA, was used to examine the effects of miR-188-5p on the sensitivity of colon cancer cells to OXA. The target of miR-188-5p was identified using a luciferase assay. Cell cycle distribution was also assessed using flow cytometry. The measurement of p21 protein expression, Hoechst 33342 staining and Annexin V/propidium iodide staining was used to evaluate apoptosis. The expression of miR-188-5p significantly increased in SW480/OXA compared with wild-type SW480 cells. The luciferase assay demonstrated that miR-188-5p inhibited Ras GTPase-activating protein 1 (RASA1; also known as p120/RasGAP) luciferase activity by binding to the 3′-untranslated region of RASA1 mRNA, suggesting that miR-188-5p could target RASA1. In addition, miR-188-5p downregulation or RASA1 overexpression promoted the chemosensitivity of SW480/OXA, as evidenced by increased apoptosis and G₁/S cell cycle arrest. Moreover, RASA1 silencing abrogated the increase in cell apoptosis induced by the miR-188-5p inhibitor. The findings of the present study suggested that miR-188-5p could enhance colon cancer cell chemosensitivity by promoting the expression of RASA1.     

上调结肠癌间充质干细胞中miR-93-5p表达抑制结肠癌细胞增殖及促进凋亡的机制探讨

目的:观察miR-93-5p上调的结肠癌间充质干细胞(colon cancer mesenchymal stem cell,CCMSC)对结肠癌细胞增殖、凋亡的影响,并初步探讨其机制.方法:以贴壁细胞培养法抽提癌组织CCMSC,从正常结肠组织抽提结肠间充质干细胞(colon mesenchymal stem cell,...     

LINC00909靶向miR-365a-5p/FGFBP1分子轴调控结肠癌细胞增殖、迁移及侵袭的实验研究

目的 探讨LINC00909对结肠癌细胞增殖、迁移、侵袭的影响及可能机制.方法 购买正常结肠上皮细胞NCM460,结肠癌细胞HCT8、Caco-2及DLD-1;用实时荧光定量PCR(RT-qPCR)及蛋白质印迹法(Western blotting)检测NCM460细胞及结肠癌细胞中LINC00909、miR-365a-...     

miR－647在结肠癌细胞中的表达及临床意义

目的：分析miR－647在结肠癌组织及细胞系中的表达情况,探讨其对结肠癌细胞增殖、迁移能力的影响及其可能的作用机理。方法：利用Real－time quantitative Polymerase Chain Reaction（qPCR）技术检测17例结肠癌患者癌及癌旁组织中miR－647的表达;利用qPCR技术检测正常人肠上皮细胞HIEC及结肠癌细胞HT－29中miR－647的表达;将miR－647 antagomir及对照分别转染至结肠癌细胞中,应用MTT实验检测细胞增殖,体外划痕实验检测细胞迁移能力,评价转染miR－647抑制剂对结肠癌细胞增殖能力和迁移能力的影响。结果：qPCR结果显示,与癌旁组织相比,miR－647在结肠癌肿瘤组织中表达明显升高;与正常人肠上皮细胞相比,人结肠癌细胞系HT－29中miR－647表达明显升高;MTT结果显示miR－647抑制剂可以显著抑制SW480和SW620细胞的增殖能力和细胞迁移能力。结论：miR－647通过促进结肠癌细胞的增殖和迁移能力,参与结肠癌的发生发展进程。     

结肠癌组织中miR-802和RAB23的表达水平及临床意义

目的:检测微小RNA-802(miR-802)和RAB23在结肠癌组织中的表达水平,探究其临床意义。方法:以本院2014年3月至2015年4月收治的结肠癌患者84例为研究对象,所有患者经诊断后行结肠癌根治术,术中取患者癌组织及癌旁正常组织。实时荧光定量PCR检测miR-802和RAB23 mRNA表达水平,免疫组化分析RAB蛋白表达水平。结果:miR-802在结肠癌组织中的表达水平显著低于癌旁正常组织,RAB23表达水平显著高于癌旁正常组织（P<0.05）;结肠癌组织中RAB23蛋白表达阳性率显著高于癌旁正常组织（P<0.05）;相关性分析表明结肠癌组织中miR-802表达水平与RAB23 mRNA表达水平呈负相关（P<0.05）;miR-802和RAB23表达水平与患者肿瘤大小和淋巴转移情况有关（P<0.05）;结肠癌患者中miR-802低表达者、RAB23阳性表达者生存率分别显著低于miR-802高表达者、RAB23阴性表达者（P<0.05）;COX分析显示miR-802和RAB23表达水平是影响结肠患者不良预后的独立危险因素（P<0.05）。结论:miR-802在结肠癌患者癌组织中表达水平显著降低,RAB23表达水平显著升高,与结肠癌患者预后有关,可为结肠癌患者预后判断提供参考。     

miR-520a在结肠癌组织中的表达及意义

目的:探讨miR-520a在结肠腺瘤性息肉、结肠癌及癌旁组织中的表达变化,并分析miR-520a在结肠癌中的表达与临床病理特征的关系。方法:采用实时荧光定量PCR（qRT-PCR）检测30例结肠腺瘤组织和55例结肠癌及其癌旁组织中miR-520a的表达,分析结肠癌中miR-520a的表达与临床病理特征之间的关系。结果:miR-520a在结肠癌组织中的表达明显低于结肠腺瘤,而结肠腺瘤中的表达明显低于癌旁组织（P＜0.05）。在结肠癌中,miR-520a的表达与肿瘤分化程度、淋巴转移、TNM分期、远处转移均显著相关（P＜0.05）。结论:miR-520a在结肠癌中低表达,提示其可能发挥抑癌基因的功能。     

结肠癌患者循环miR-630的表达及其临床意义

目的:探讨结肠癌患者外周血中microRNA-630(miR-630)的表达,进一步分析其表达对评估结肠癌进展及预后的价值。方法:使用实时荧光定量PCR（Real time PCR）检测55例结肠癌及21例正常对照者外周血中miR-630的表达,采用统计学方法分析miR-630在结肠癌外周血中的表达及其与肿瘤进展和预后的关系。结果:分析Real time PCR检测结果发现,结肠癌患者外周血中miR-630的表达显著高于正常对照（P<0.05）,外周血中miR-630表达与结肠癌的侵袭、转移和分期显著相关（P<0.05）,单因素和多因素生存分析发现外周血中miR-630的表达上调与结肠癌预后不良显著相关（P<0.05）。结论:结肠癌患者循环miR-630表达显著上调,与结肠癌侵袭和转移密切相关,并可作为结肠癌的独立预后标记分子。     

TRAP1在结肠癌组织中的表达与病理特征和患者预后的关系及其可能的机制

目的：探讨肿瘤坏死因子受体相关蛋白1(TRAP1)在结肠癌组织和细胞中的表达及其与临床病理特征和患者预后的关系和相关分子机制。方法：通过TCGA和GEO数据全面分析TRAP1在结肠癌中的表达及其与临床病理特征和患者预后的关系，选取2020年10月至2021年03月间在山西医科大学第一医院手术切除的10例结肠癌组织及相应癌旁组织标本，用IHC染色法检测中国人结肠癌组织中TRAP1的表达进行验证，运行R包（survival和survminer）进行Kaplan-Meier生存分析；在线分析TRAP1蛋白的信号肽及穿膜结构域，通过基因富集分析软件进行GO分析和KEGG分析。培养结肠癌SW480和SW620细胞，将si-NC和si-TRAP1转染结肠癌细胞，实验分为空白对照组、si-NC组和si-TRAP1组，采用qPCR法检测转染后各组结肠癌细胞中TRAP1的表达，FCM检测转染后各组细胞的细胞周期和凋亡情况。结果：与癌旁组织比较，TRAP1在结肠癌组织中呈高表达（P<0.01）,TRAP1表达水平与淋巴结转移有关联（P<0.05）,TRAP1高表达组结肠癌患者5年OS率较低（P...     

结肠癌T-cadherin表达水平分析与5-Aza-CdR对其表达和结肠癌细胞增殖、侵袭及凋亡的影响

背景与目的：结肠癌是临床常见恶性肿瘤,探讨抑癌蛋白T-cadherin在结肠癌中的表达情况及其与患者临床病理学特征的关系,并分析5-Aza-CdR对T-cadherin表达和结肠癌细胞增殖、侵袭及凋亡的影响。方法：收集福建医科大学附属第二医院2015—2016年40例手术切除的结肠癌组织及癌旁组织新鲜样本,并经过病理学检查验证,分别提取总RNA和总蛋白质。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）分析T-cadherin mRNA的表达,采用蛋白质印迹法（Western blot）分析T-cadherin蛋白水平,并分析T-cadherin的mRNA表达变化与患者临床病理学特征的关系。进一步以人结肠癌细胞系HT-29为研究对象,采用甲基化抑制剂5-Aza-CdR处理HT-29细胞,分别采用RTFQ-PCR和Western blot分析T-cadherin表达变化,采用细胞计数试剂盒（cell counting kit-8,CCK-8）分析细胞增殖,采用Transwell实验验证细胞侵袭能力,采用流式细胞术分析细胞凋亡。结果：T-cadherin在结肠癌组织的蛋白水平和mRNA表达均明显低于癌旁组织,其mRNA表达与淋巴结转移（F=5.316,P=0.009 3）和分化程度（F=5.807,P=0.006 4）明显相关,而与其他病理变量（包括性别、年龄、肿瘤大小和肿瘤浸润深度）无明显相关。药物5-Aza-CdR可以显著上调HT-29细胞中T-cadherin的表达水平,抑制HT-29细胞增殖、侵袭并促进细胞凋亡。结论：T-cadherin表达可能与结肠癌恶性程度密切相关,药物5-Aza-CdR处理可上调结肠癌细胞T-cadherin的表达,并抑制结肠癌细胞的增殖、迁移,促进结肠癌细胞凋亡,提示T-cadherin可能是结肠癌患者使用甲基化抑制剂5-Aza-CdR治疗的靶点。     

TMIGD1在结肠癌中的表达及对结肠癌细胞增殖和侵袭的影响

目的 检测TMIGD1在结肠癌及其相应的癌旁正常结肠上皮组织中的表达情况及过表达TMIGD1对结肠癌细胞生物学行为的影响,并探讨潜在分子机制.方法 通过生物信息学数据库和免疫组织化学染色检测TMIGD1在结肠癌与癌旁正常组织中的表达情况,并分析其与不同临床病理因素之间的关系.对结肠癌细胞系SW480通过慢病毒包装系统使...     

miR-191 promotes tumorigenesis of human colorectal cancer through targeting C/EBPβ

MicroRNA-191 (miR-191), a small non-coding RNA, is involved in disease development and cancer diagnosis and prognosis. However, how miR-191 functions in colorectal cancer remains largely unclear. In this study, we show that miR-191 is highly expressed in colon tumor tissues, and that inhibition of miR-191 leads to decreased cell growth, proliferation and tumorigenicity in a xenograft model. Overexpression of miR-191 in colorectal cancer cell lines alters cell cycle progression and cell resistance to 5-Fu induced cell apoptosis. Mechanistic studies demonstrated that miR-191 directly binds to the 3′UTR of the C/EBPβ mRNA and mediates a decrease in the mRNA and protein expression of C/EBPβ. We further showed that C/EBPβ induces growth arrest in a colorectal cancer cell line and that its expression is negatively correlated with the miR-191 level in patient samples. Our findings suggest that miR-191 may be a potential gene therapy target for the treatment of colorectal cancer.     

Expression and clinical significance of miR-17-5p in tumor tissues of patients with colorectal cancer

BackgroundAt present, there is a lack of novel biomarkers for the early diagnosis and targeted therapy of colorectal cancer (CRC). The current study investigated the expression of miR-17-5p in colorectal tumors and adjacent tissues, and examined the effects of miR-17-5p on the cellular growth of the colon cancer cell line HCT-116.MethodsA total of 30 paired tissue specimens were obtained from patients with CRC who were admitted to the Department of General Surgery V of the First Affiliated Hospital, Gannan Medical College between December 2019 and January 2021. The clinical information for the corresponding patients was collated. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was performed to detect the expression of miR-17-5p in tumor tissues and the paracancerous tissues. The relationship between miR-17-5p and clinicopathology was analyzed. The CRC cell line HCT-116 was transfected with miR-17-5p mimics and inhibitors using liposomes. The effects of miR-17-5p on cell proliferation, invasion and migration, and apoptosis were determined using colon formation assays, transwell and scratch assays, and flow cytometry, respectively. The effects of miR-17-5p on CD44 (homing cell adhesion molecule), MMP2 (matrix metalloproteinase 2), BCL2 (B-cell lymphoma-2), BAX (BCL2-associated X), and E-cadherin protein and gene expression were investigated using Western blot and RT-qPCR, respectively.ResultsMiR-17-5p expression was higher in tumor tissues compared to the normal adjacent tissues. While miR-17-5p expression levels were unrelated to age nor gender, they were related to the degree of differentiation, stage, lymph node metastasis, and tumor size in patients with CRC. Upregulation of miR-17-5p promoted CRC cell proliferation, metastasis, and invasion, while inhibiting apoptosis. However, downregulation of miR-17-5p had the opposite effect. Furthermore, miR-17-5p upregulation increased E-cadherin, CD44, MMP2, and BCL2 expression while decreasing BAX expression, whereas miR-17-5p downregulation had the opposite effect.ConclusionsMiR-17-5p is highly expressed in tumor tissues and correlates with increased proliferation, invasion, and migration, as well as reduced apoptosis in HCT-116 cells. Indeed, miR-17-5p may be a potential indicator for the early diagnosis of CRC and may guide clinical targeted therapy.     

肠癌组织中miR-141的表达及其对HCT116细胞增殖和迁移的影响

目的:探讨miR-141在肠癌组织中的表达情况以及其对HCT116细胞生物学功能的影响及其作用机制。方法:选取2016年05月至2018年05月在中国医科大学附属第一人民医院手术切除的30对肠癌组织以及癌旁组织进行miRNA芯片筛查。逆转录定量聚合酶链反应分析其中异常表达miRNA情况,评估miR-141的表达与肿瘤相关信息的相关性。通过TargetScan软件分析miR-141可能靶向的蛋白,在HCT116细胞中通过荧光素酶报告基因实验检测miR-141对DEK蛋白的靶向作用。HCT116细胞中分别过度表达和沉默表达miR-141后,通过MTT实验检测细胞增殖情况,Transwell实验检测细胞的迁移情况。结果:芯片分析和逆转录定量聚合酶链反应指出miR-141在肠癌组织中表达低于癌旁组织,miR-141与肿瘤的进程具有相关性,TargetScan软件指出miR-141可以靶向作用于DEK,荧光素酶报告基因实验印证了miR-141对DEK的靶向作用。MTT实验指出miR-141过度表达显著抑制细胞增殖,miR-141沉默表达显著促进细胞增殖,与对照组相比,差异有统计学意义（P＜0.05）。Transwell实验显示,miR-141过度表达可以抑制HCT116细胞的迁移,miR-141沉默表达后HCT116细胞的迁移得到促进。结论:miR-141通过靶向DEK蛋白能够抑制HCT116细胞的增殖和迁移。     

微小RNA-218在胃癌中的表达与作用

目的 检测微小RNA-218(miR-218)在胃癌组织中的表达水平及其功能,探讨miR-218在胃癌发生、发展中的作用和意义.方法 采用发夹状引物的逆转录聚合酶链反应(RT-PCR)技术,检测20例非贲门胃癌组织和正常胃黏膜组织中miR-218的表达水平.通过转染不同的外源性分子,改变胃上皮AGS细胞miR-218的表达水平,检测miR-218对细胞增殖、凋亡、细胞周期和侵袭能力的影响.结果 与正常胃黏膜组织相比,miR-218在胃癌组织中呈低表达(P＜0.01),其比值平均为0.19(范围0.01～0.70).流式细胞技术检测结果显示,转染miR-218前体的AGS细胞凋亡率(21.6%)明显增加,与转染阴性参照(10.4%)的AGS细胞差异有统计学意义(P＜0.05).与转染阴性参照比较,转染miR-218前体可明显抑制细胞增殖(P＜0.01).Transwell试验证实,AGS细胞在转染miR-218前体后,细胞侵袭能力明显减弱(P＜0.05).结论 在胃癌组织中,miR-218的表达降低,miR-218具有促进细胞凋亡、抑制细胞增殖和侵袭的功能.