miR-551b-3p在胃癌组织中低表达并抑制胃癌细胞系HGC-27的增殖、迁移和侵袭

目的探讨miR-551b-3p在胃癌中的表达变化情况,及其对胃癌细胞功能的影响。方法用real-time PCR的方法检测60例胃癌组织及其对应的癌旁组织中miR-551b-3p的表达量,使用miR-551b-3p模拟物(miR-551b-3p mimic)转染胃癌细胞系HGC-27,CCK-8法检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell法检测细胞侵袭。结果 1)miR-551b-3p在胃癌癌症组织中的表达明显低于癌旁组织(P0.05)。2)过表达miR-551b-3p mimic可以明显减弱胃癌细胞系HGC-27的增殖、迁移和侵袭能力。结论 miR-551b-3p在胃癌组织中低表达,并且抑制胃癌细胞系HGC-27的增殖、迁移和侵袭,可能与胃癌发生发展密切相关。     

Mi R-31在胃癌组织中低表达并抑制HGC-27细胞增殖、迁移与侵袭

目的研究miR-31在胃癌患者中的表达,探讨miR-31对胃癌HGC-27细胞增殖、侵袭和转移的影响。方法收集胃癌患者标本39例,同时收取相应的癌旁组织20例。通过荧光定量PCR方法检测各组织中miR-31的表达水平。以脂质体为载体将miR-31模拟物转染至HGC-27细胞,CCK-8方法检测细胞增殖能力变化,Transwell实验检测HGC-27细胞侵袭和迁移能力变化。结果 miR-31在胃癌组织中的表达水平明显低于癌旁组织,P0.01。过表达miR-31能够显著抑制HGC-27细胞的增殖,显著抑制HGC-27细胞的侵袭和迁移能力(P0.01)。结论 MiR-31在胃癌中低表达,并抑制胃癌细胞系HGC-27的增殖、迁移和侵袭。     

miR-760对胃癌细胞系MGC-803增殖、迁移和侵袭的影响

目的探讨miR-760对胃癌细胞系MGC-803增殖、迁移和侵袭的影响。方法 Real-time PCR分析50例胃癌组织(C)及其癌旁(N)中miR-760的表达水平;用pc DNA3.1载体构建过表达miR-760的重组质粒(pc DNA-miR-760),实现miR-760在MGC-803细胞中的过表达;分别用CCK-8法、Transwell和划痕实验检测细胞增殖、侵袭和迁移能力。结果与癌旁对照组相比,36例(72%)胃癌组织中出现miR-760的表达下调;过表达miR-760能显著抑制MGC-803细胞的迁移和侵袭能力(P0.05),但对其增殖影响不大。结论 miR-760的表达下调可能与胃癌的进展有关,过表达miR-760可以抑制胃癌MGC-803细胞的迁移和侵袭。     

miR-363-3p在胃癌组织中的表达及其对细胞系HGC-27增殖、迁移与侵袭功能的影响

目的探讨miR-363-3p在胃癌及癌旁样本中的表达差异,并分析其在胃癌细胞系中的功能。方法使用realtime PCR检测59例胃癌组织及对应癌旁组织中miR-363-3p的表达差异;使用miR-363-3p模拟物(miR-363-3pmimic)实现miRNA在胃癌细胞系HGC-27中的过表达,经增殖实验、划痕实验和Transwell实验,检测miR-363-3p对HGC-27细胞功能的影响。结果 miR-363-3p抑制胃癌细胞系HGC-27细胞增殖(P0.05,P0.01),抑制胃癌细胞系HGC-27细胞迁移(P0.001),miR-363-3p对抑制胃癌细胞系细胞的侵袭(P0.05)。结论 miR-363-3p在胃癌发生中可能起到抑癌作用,并有可能成为对胃癌治疗的一个新靶点。     

DADS通过miR-222抑制人胃癌细胞系的增殖与侵袭

目的通过miRNA途径研究二烯丙基二硫(DADS)的抑瘤机制,以进一步阐明DADS抑制胃癌细胞增殖与转移的分子机制。方法将胃癌细胞系MGC-803细胞分为DADS处理组、miR-222模拟物组、miR-222抑制物组和阴性对照组;分别采用0、25、50、100、200和400μmol/L DADS处理MGC-803细胞;分别将miR-222模拟物、miR-222抑制物和scramble转染MGC-803细胞。qRT-PCR检测MGC-803细胞中miR-222表达;MTT法和Transwell侵袭实验检测MGC-803细胞增殖与侵袭能力;蛋白质印迹试验检测MGC-803细胞中TIMP3蛋白表达。结果 DADS可呈剂量依赖性下调MGC-803细胞中miR-222表达(P0.05);DADS能抑制MGC-803细胞的增殖与侵袭,外源高表达miR-222能促进MGC-803细胞的增殖与侵袭,而miR-222抑制物与DADS共同处理,MGC-803细胞的增殖与侵袭抑制作用最为显著(P0.05);DADS可下调MGC-803细胞中TIMP3蛋白的表达,外源高表达miR-222能上调MGC-803细胞中TIMP3蛋白的表达,而miR-222抑制物与DADS共同处理,MGC-803细胞中TIMP3蛋白的表达下调最为显著(P0.05)。结论 DADS通过下调miR-222的表达,靶向TIMP3抑制胃癌细胞的增殖与侵袭。     

MicroRNA-29b重组慢病毒表达载体的构建及其抑制胃癌细胞系的迁移和增殖

目的构建has-miR-29b重组慢病毒的表达载体,并观察其对胃癌细胞系迁移和增殖能力的影响。方法将PCR扩增的miR-29b前体序列和pMIR载体经双酶切后连接产生pMIR-miR-29b慢病毒表达载体。pMIR-miR-29b、Packaging Plasmid(s)和pVSV-G质粒共转染包装细胞系293TN,包装产生慢病毒。使用收获的慢病毒颗粒感染胃癌细胞系HGC-27,72 h后利用real-time PCR检测miR-29b在HGC-27内的表达水平,Western blot检测其靶基因MCL-1的表达。划痕实验和细胞增殖实验分别观察在HGC-27细胞中过表达miR-29b后细胞迁移能力及增殖能力的变化。结果成功构建了miR-29b重组慢病毒的表达载体,感染胃癌细胞HGC-27后,能够成功过表达miR-29b。在HGC-27细胞中过表达miR-29b,其靶基因MCL-1的表达明显被抑制,并且Lenti-29b组与对照组Untreat组和Lenti-vector组相比,Lenti-29b组的迁移能力显著降低(P<0.01);Lenti-29b组与Lenti-vector组相比,Lenti-29b组在...     

MiR-33a在胃癌组织中低表达并抑制HGC-27细胞增殖、迁移与侵袭

目的探讨miR-33a在胃癌患者中的表达,探讨miR-33a对胃癌HGC-27细胞增殖、侵袭和转移的影响。方法收集胃癌患者标本39例,同时收取相应的癌旁组织20例。通过荧光定量PCR方法检测各组织中miR-33a的表达水平。以脂质体为载体将miR-33a模拟物转染至HGC-27细胞,CCK-8方法检测细胞增殖能力变化,Transwell实验检测HGC-27细胞侵袭和迁移能力变化。结果 miR-33a在胃癌组织中的表达水平明显低于癌旁组织,过表达miR-33a能够显著抑制HGC-27细胞的增殖,显著抑制HGC-27细胞的侵袭和迁移能力。结论 miR-33a可能对胃癌有治疗作用。     

miR-433-3p靶向HP1BP3抑制胃癌细胞的增殖、迁移和侵袭能力

目的：探究微小RNA-433-3p(miR-433-3p)对胃癌细胞增殖、迁移和侵袭的影响，以及miR-433-3p靶向异染色质蛋白1结合蛋白3(HP1BP3)对胃癌细胞的调控作用。方法：将人胃癌细胞系HGC-27和AGS分为阴性对照（NC）mimic组、miR-433-3p mimic组、NC siRNA (si-NC)组和HP1BP3 siRNA (si-HP1BP3)组，分别转染NC mimic、miR-433-3p mimic、si-NC和si-HP1BP3。RT-qPCR检测miR-433-3p在HGC-27和AGS细胞中的表达；CCK-8法和细胞集落形成实验检测细胞增殖；划痕愈合实验检测细胞迁移；Traswell实验检测细胞侵袭；Western blot检测HP1BP3蛋白表达。结果：与NC mimic组相比，miR-433-3p mimic组HGC-27和AGS细胞中miR-433-3p表达升高。过表达miR-433-3p抑制HGC-27和AGS细胞的增殖、迁移和侵袭，抑制HP1BP3蛋白表达；敲减HP1BP3抑制HGC-27和AGS细胞的增殖和迁移。结论：miR-433...     

miR-216a-5p通过抑制HMGB1降低胃癌细胞系MGC-803的增殖和迁移

目的探讨miR-216a-5p对胃癌细胞增殖和迁移的影响及其分子机制。方法用Western blot检测胃癌和癌旁组织中高迁移率族蛋白1(HMGB1)的表达,RT-qPCR检测miR-216a-5p的表达。用人工合成的miR-216a-5p模拟物和其抑制物转染胃癌细胞系MGC-803。MTT检测细胞增殖,划痕法检测细胞迁移。双荧光素酶报告实验和Western blot检测miR-216a-5p和HMGB1之间的靶向关系。结果与癌旁组织相比,HMGB1在胃癌组织中高表达,miR-216a-5p在胃癌组织中低表达(P0.05)。miR-216a-5p模拟物上调胃癌细胞miR-216a-5p的表达水平,抑制细胞的增殖和迁移;miR-216a-5p抑制物下调miR-216a-5p的表达水平,促进胃癌细胞的增殖和迁移(P0.05)。miR-216a-5p可作用于HMGB1的3′UTR并抑制其表达。结论 miR-216a-5p通过靶向抑制HMGB1的表达降低胃癌细胞系MGC-803的增殖和迁移。     

WTAP基因在胃癌组织中低表达

目的探讨WTAP基因在胃癌组织样本中的表达变化情况,并检测其对胃癌细胞表型的影响。方法用RTq PCR的方法检测45例胃癌组织及其对应的癌旁组织中WTAP mRNA的表达量,并结合临床病理资料进行分析;使用RNA干扰方法敲低内源性WTAP在胃癌细胞系MGC-803中的表达,再通过CCK-8实验、划痕迁移实验和transwell实验,检测WTAP表达降低后对MGC-803细胞功能的影响。结果 WTAP mRNA在胃癌癌症组织中的表达明显低于癌旁组织(P0.05),且其表达与胃癌样本的临床分期密切相关(P0.05);在MGC-803细胞系中敲低WTAP,可促进胃癌细胞增殖、促进细胞迁移和侵袭。结论 WTAP在胃癌组织中低表达,可能与其发生发展有关,有望成为胃癌诊治的靶标。     

特异性微小RNA抑制剂对胃癌细胞增殖的影响

目的： 探讨特异性微小RNA抑制剂对胃癌细胞增殖的影响。方法: 设计并合成4种微小RNA（miR-17、miR-21、miR-106a和miR-421）的2’-甲氧修饰的RNA寡核苷酸（微小RNA抑制剂）,用脂质体分别转染到SGC-7901和MGC-803胃癌细胞,然后用实时定量逆转录-聚合酶链反应技术检测微小RNA的表达情况,最后用MTT方法检测胃癌细胞的增殖情况。结果: 4种微小RNA抑制剂均有效抑制了SGC-7901和MGC-803细胞中相应微小RNA的表达;除miR-21抑制剂未见抑制胃癌细胞的增殖外,其它3种微小RNA抑制剂均以剂量依赖方式抑制胃癌细胞的增殖且对SGC-7901的效果优于MGC-803。结论: 微小RNA的特异性抑制剂能有效抑制胃癌细胞的增殖,采用这种技术为研究胃癌的发病机制提供了新方法。     

致癌性微小RNA-106a对正常胃黏膜上皮细胞和胃癌细胞生长的影响

目的: 探讨微小RNA-106a(miR-106a)是否具有致癌特性。方法: 用脂质体把miR-106a类似物转染到正常胃黏膜上皮细胞GES-1中,然后用MTT方法检测该细胞的增殖情况。用脂质体把miR-106a抑制剂转染到胃癌细胞MGC-803和SGC-7901中,然后分别用流式细胞术和Western印迹方法检测细胞周期分布和细胞周期相关蛋白表达的变化;最后观察裸鼠胃癌移植瘤的生长情况。结果: miR-106a类似物以剂量依赖方式促进正常胃黏膜上皮细胞的增殖。miR-106a抑制剂通过抑制细胞周期素依赖的蛋白激酶(CDK)1和CDK2的表达阻滞MGC-803细胞于G₀/G₁期和G₂/M期,而通过抑制CDK1的表达阻滞SGC-7901于G₂/M期。动物实验结果显示,miR-106a抑制剂以剂量依赖方式抑制肿瘤的生长。结论: miR-106a可能参与了胃癌的发生过程。     

MiR-125b promotes migration and invasion by targeting the vitamin D receptor in renal cell carcinoma

Purpose: To investigate the expression of miR-125b and vitamin D receptor (VDR) in renal cell carcinoma (RCC) and assess the biological function of miR-125b in RCC.Methods: We used quantitative real-time polymerase chain reaction (RT-PCR) to detect the expression of nucleic acids and western blotting to analyze the protein abundance in RCC cell lines. MiR-125b mimic and inhibitor were employed to investigate the function and behavior of miR-125b in RCC cell lines. The relationship between miR-125 and VDR was verified using luciferase assays.Results: Overexpression of miR-125b promoted migration and invasion and prevent cell apoptosis in ACHN cells. In contrast, miR-125b deficiency suppressed migration and invasion and induced cell apoptosis in 786-O cells. Luciferase assays indicated the interaction between miR-125b and VDR. In collected samples, miR-125b was significantly higher in RCC tissues and negatively correlated to VDR (r=-0.444, p=0.04).Conclusion: MiR-125b displays an oncogene profile in RCC, patients with high expression of miR-125b should be a more frequent follow-up. MiR-125B may be a potential therapeutic target for RCC.     

Long non-coding RNA ncRuPAR regulates gastric cancer cell proliferation and apoptosis via phosphoinositide 3-kinase/protein kinase B signaling

Objective: To determine the effect and mechanism of the long non-coding RNA (lncRNA) ncRuPAR (non-protein coding RNA, upstream of coagulation factor II thrombin receptor [F2R]/protease-activated receptor-1 [PAR-1]) in human gastric cancer.Methods: HGC-27-ncRuPAR overexpression and MGC-803-ncRuPAR-RNAi knockdown gastric cancer cell lines were established. We assessed the effect of ncRuPAR on cell proliferation, apoptosis, migration, and invasion using Cell Counting Kit 8, flow cytometry, scratch and transwell assays, respectively. Differentially expressed genes in HGC-27-ncRuPAR overexpression and HGC-27-empty vector cell lines were identified using Affymetrix GeneChip microarray analysis. Ingenuity Pathway Analysis (IPA) of the microarray results was subsequently conducted to identify ncRuPAR-enriched pathways, followed by validation using real time-quantitative PCR (RT-qPCR). As one of the top enriched pathways, phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway was further examined by western blotting to determine its role in ncRuPAR-mediated regulation of gastric cancer pathogenesis.Results: ncRuPAR inhibited human gastric cancer cell proliferation and induced G1/S phase arrest and apoptosis, but did not affect migration or invasion in vitro. Overexpression of ncRuPAR in vitro was found to inhibit its known target PAR-1, as well as PI3K/Akt signaling. The downstream targets of PI3K/Akt, cyclin D1 was downregulated, but there was no change in expression level of B-cell lymphoma 2 (Bcl-2).Conclusions: We showed that lncRNA-ncRuPAR could inhibit tumor cell proliferation and promote apoptosis of human gastric cancer cells, potentially by inhibiting PAR-1, PI3K/Akt signaling, and cyclin D1. The results suggest a potential role for lncRNAs as key regulatory hubs in GC progression.     

FTO在胃癌组织中的表达降低及其对细胞系MGC-803功能的影响

目的探讨RNA甲基化相关蛋白FTO在胃癌中的表达变化情况,及其对胃癌细胞功能的影响。方法用realtime PCR检测54例胃癌组织及对应癌旁组织中FTO的表达;用质粒(p CMV6-FTO)实现FTO在胃癌细胞系MGC-803中的过表达;通过增殖实验、划痕试验和Transwell实验,检测FTO对MGC-803细胞功能的影响。结果 FTO mRNA在胃癌组织的表达水平显著低于癌旁组织(P0.05),且其表达与胃癌样本的临床分期密切相关(P0.05);在MGC-803细胞系中过表达FTO,可抑制胃癌细胞增殖(P0.05)、抑制细胞迁移(P0.05)和侵袭(P0.05)。结论 FTO在胃癌组织中低表达,并且抑制胃癌细胞系的增殖、迁移和侵袭,可能与胃癌发生发展密切相关。     

Downregulation of TRIM27 suppresses gastric cancer cell proliferation via inhibition of the Hippo-BIRC5 pathway

Although tripartite motif containing 27 (TRIM27) protein has been implicated in the progression of many cancer types, its role in gastric cancer (GC) remains poorly understood. Given that TRIM27 may be associated with the baculoviral inhibitor of apoptosis repeat containing 5 (BIRC5) gene, which is downstream of the Hippo pathway, we clarified their relationship in GC progression. In vitro cultures of 7 GC cell lines, 92 GC patient tumor samples and 46 normal clinical samples were used to examine the influence of changes in TRIM27 expression, which was assessed by quantitative PCR, immunohistochemistry, western blot analysis, and cell viability assays. We found that TRIM27 overexpression was correlated with tumor size, depth of invasion, and poor GC prognosis, while TRIM27 small interfering RNA knockdown inhibited cell proliferation and colony formation, induced apoptosis, and increased sensitivity towards 5-fluorouracil treatment in MGC-803 and HGC-27 GC cell lines. Notably, TRIM27 downregulation resulted in BIRC5 suppression via large tumor suppressor kinase 2 (LATS2) upregulation and subsequent Yes-associated protein 1 (YAP1) inhibition in MGC-803 and HGC-27 GC cell lines. In conclusion, our findings revealed the positive correlation between TRIM27 and GC progression through mediation of the Hippo-BIRC5 axis in GC.     

MicroRNA-34a inhibits tumor invasion and metastasis in gastric cancer by targeting Tgif2

To study how miR-34a acts as a tumor suppressor in inhibiting the invasion and metastasis of the gastric cancer cells. First, real-time polymerase chain reaction (PCR) and western blot analysis were used to analyze the expression of miR-34a and Tgif2 in gastric cancer tissues and the adjacent normal tissues. Next, gastric cancer cells were transfected with miR-34a mimic and Tgif2 siRNA, respectively. After transfection, real-time PCR and western blot analysis were used to detect the relative Tgif2 expression level. Cell proliferation was monitored by the colorimetric water-soluble tetrazolium salt and apoptosis analysis was performed with Annexin-V-FITC Apoptosis Detection Kit I. The expression of miR-34a in the adjacent non-tumor tissues was higher than that in gastric cancer tissues, but Tgif2 was opposite. In gastric cancer cells transfected with miR-34a mimic/Tgif siRNA, Tgif2 expression was remarkably down-regulated. Cells transfected with miR-34a mimic/Tgif2 siRNA grew more slowly than the control groups. The percentage of apoptotic cells in gastric cancer cells transfected with miR-34a mimic/Tgif siRNA was much higher compared to the controls. Therefore, we concluded that miR-34a could inhibit tumor invasion and metastasis in gastric cancer by targeting Tgif2 and may be a novel therapeutic candidate for gastric cancer.     

MicroRNA-301b-3p accelerates the growth of gastric cancer cells by targeting zinc finger and BTB domain containing 4

MicroRNAs (miRNAs) have been found to be aberrantly expressed and exert essential roles in the tumorigenesis and progression of gastric cancer (GC). miR-301b-3p has been recognized as a cancer-related miRNA in lung cancer, bladder cancer and hepatocellular carcinoma. However, the function of miR-301b-3p in GC progression and its underlying mechanism have not been studied yet. In this study, we found that miR-301b-3p expression was up-regulated in GC tissues compared to adjacent noncancerous tissues. Furthermore, the elevated levels of miR-301b-3p were detected in GC cell lines (SGC-7901, AGS, MKN-45 and MGC-803) as compared with GES-1 cells. Interestingly, GC tissues from patients with tumor size ≥ 5 cm and advanced tumor stages showed obvious higher levels of miR-301b-3p compared to matched controls. Functionally, miR-301b-3p knockdown prominently inhibited cell proliferation, and induced cell cycle arrest at G1 phase and apoptosis in MGC-803 cells. Meanwhile, ectopic expression of miR-301b-3p conversely regulated these biological behaviors of MKN-45 cells. Next, we found that miR-301b-3p knockdown increased, whereas miR-301b-3p overexpression reduced the expression of zinc finger and BTB domain containing 4 (ZBTB4) in GC cells. Accordingly, luciferase reporter assay identified ZBTB4 as a direct target of miR-301b-3p. ZBTB4 overexpression markedly restrained the growth of MGC-803 cells. More importantly, ZBTB4 silencing partially reversed miR-301b-3p knockdown-induced tumor suppressive effects on MGC-803 cells. In conclusion, we firstly revealed that miR-301-3p was highly expressed in GC and contributed to tumor progression via attenuating ZBTB4, which might provide a novel molecular-targeted strategy for GC treatment.     

MicroRNA-101 regulates the viability and invasion of cervical cancer cells

Background: Cervical cancer has the second highest morbidity and mortality rates of any malignancy in women worldwide, and it is one of the leading causes of death in Uygur women in Xinjiang China. MicroRNAs are involved in cancer development and progression. Previously, we found that miR-101 is significantly down-regulated in cervical cancer tissues from Uyghur women. The underlying pathophysiology and relevance to tumorigenesis of miR-101 is still largely unknown. The purpose of this study was to elucidate the molecular mechanisms of miR-101 regulation of cervical cancer cell viability and invasion. Materials and methods: The expression of miR-101 in cervical cancer cell line (SiHa) was detected by real-time PCR. A miR-101 mimic was overexpressed in SiHa cells, and MTT assays were performed to determine the impact on cell proliferation. Cell would heal assays and flow cytometry were used to detect migratory ability and cellular apoptosis, respectively. Immunohistochemistry was performed to assess protein expression of the miR-101 target gene COX-2. Results: MiR-101 was endogenously expressed in SiHa cells, and alterations in its expression had profound effects on cellular migration and invasion efficiency. Overexpression of miR-101 decreased proliferation in the MTT assay (the mimics at 490 nm absorbance is lower 60% than normal, and decreased cellular motility in the cell would healing assay (transfected: 37 ± 2 m, pre-transfected 184 ± 2 m). Apoptosis rate was significantly higher with overexpression of miR-101 relative to control (transfected: 76.6%, pre-transfected: 3.5%) (P < 0.05). The expression of Cox-2 was decreased in transfected cells. Conclusions: MiR-101 likely acts as a tumor suppressor in cervical cancer. Overexpression of miR-101 decreased expression of its target gene Cox-2 and inhibited proliferation and invasion, and promoted apoptosis to suppress tumorigenicity. MiR-101 is a promising new target for the development of therapeutic strategies for the clinical treatment of cervical cancer.