Emodin Inhibits Colon Cancer Cell Invasion and Migration by Suppressing Epithelial–Mesenchymal Transition via the Wnt/β-Catenin Pathway

Colon cancer (CC) is the third most common cancer worldwide. Emodin is an anthraquinone-active substance that has the ability to affect tumor progression. Our study aims to explore the effects and the relevant mechanism of emodin on the invasion and migration of CC in vitro and in vivo. In our study, we found that emodin inhibited the invasion and migration abilities of RKO cells and decreased the expression of matrix metalloproteinase-7 (MMP-7), MMP-9, and vascular endothelial growth factor (VEGF) in a dose-dependent manner. Further research suggested that emodin inhibited EMT by increasing the mRNA level of E-cadherin and decreasing the expression of N-cadherin, Snail, and b-catenin. Emodin also significantly inhibited the activation of the Wnt/b-catenin signaling pathway by downregulating the expression of related downstream target genes, including TCF4, cyclin D1, and c-Myc. A Wnt/b-catenin signaling pathway agonist abolished the effect of emodin on EMT and cell mobility, suggesting that emodin exerted its regulating role through the Wnt/b-catenin pathway. The CC xenograft model was established to study the antitumor efficiency of emodin in vivo. The in vivo study further demonstrated that emodin (40 mg/kg) suppressed tumor growth by inhibiting EMT via the Wnt/b-catenin signaling pathway in vivo. Taken together, we suggest that emodin inhibits the invasion and migration of CC cells in vitro and in vivo by blocking EMT, which is related with the inhibition of the Wnt/b-catenin signaling pathway.     

Emodin Inhibits Colon Cancer Cell Invasion and Migration by Suppressing Epithelial–Mesenchymal Transition via the Wnt/b-Catenin Pathway

Colon cancer (CC) is the third most common cancer worldwide. Emodin is an anthraquinone-active substance that has the ability to affect tumor progression. Our study aims to explore the effects and the relevant mechanism of emodin on the invasion and migration of CC in vitro and in vivo. In our study, we found that emodin inhibited the invasion and migration abilities of RKO cells and decreased the expression of matrix metalloproteinase-7 (MMP-7), MMP-9, and vascular endothelial growth factor (VEGF) in a dose-dependent manner. Further research suggested that emodin inhibited EMT by increasing the mRNA level of E-cadherin and decreasing the expression of N-cadherin, Snail, and b-catenin. Emodin also significantly inhibited the activation of the Wnt/b-catenin signaling pathway by downregulating the expression of related downstream target genes, including TCF4, cyclin D1, and c-Myc. A Wnt/b-catenin signaling pathway agonist abolished the effect of emodin on EMT and cell mobility, suggesting that emodin exerted its regulating role through the Wnt/b-catenin pathway. The CC xenograft model was established to study the antitumor efficiency of emodin in vivo. The in vivo study further demonstrated that emodin (40 mg/kg) suppressed tumor growth by inhibiting EMT via the Wnt/b-catenin signaling pathway in vivo. Taken together, we suggest that emodin inhibits the invasion and migration of CC cells in vitro and in vivo by blocking EMT, which is related with the inhibition of the Wnt/b-catenin signaling pathway     

Knockdown of Long Noncoding RNA CAT104 Inhibits the Proliferation,Migration, and Invasion of Human Osteosarcoma Cells by Regulating MicroRNA-381

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. This study aimed to explore the effects of long noncoding RNA CAT104 and microRNA-381 (miR-381) on osteosarcoma cell proliferation, migration, invasion, and apoptosis, as well as the underlying potential mechanism. We found that CAT104 was highly expressed in osteosarcoma MG63 and OS-732 cells. Knockdown of CAT104 significantly inhibited OS-732 cell proliferation, migration, and invasion, but promoted cell apoptosis. CAT104 regulated the expression of miR-381, and miR-381 participated in the effects of CAT104 on OS-732 cells. Zinc finger E-box-binding homeobox 1 (ZEB1) was a direct target gene of miR-381, which was involved in the regulatory roles of miR-381 in OS-732 cell proliferation, migration, invasion, and apoptosis, as well as c-Jun N-terminal kinase (JNK) and Wnt/ -catenin pathways. In conclusion, our research verified that suppression of CAT104 exerted significant inhibitory effects on osteosarcoma cell proliferation, migration, and invasion by regulating the expression of miR-381 and downstream ZEB1, as well as JNK and Wnt/ -catenin pathways.     

IWR-1对人肝癌细胞株Hep3B的增殖及Wnt/β-catenin信号通路的抑制作用

目的 探讨IWR-1对人肝癌细胞株Hep3B细胞增殖的影响及可能的机制。方法 用不同浓度IWR-1（2、4、8、16μmol/L）处理Hep3B细胞,CCK-8法检测细胞生长抑制率;流式细胞仪检测细胞周期变化及细胞凋亡率;Western blot法检测细胞中蛋白表达的变化,实时定量RT-PCR检测β-catenin和c-myc mRNA表达的变化。结果 IWR-1对人肝癌Hep3B细胞的生长抑制作用呈现剂量和时间依赖性（P<0.01）。流式细胞术结果显示,Hep3B细胞的细胞周期呈现明显的G0/G1期阻滞,且细胞凋亡率明显升高（P<0.01）,呈现剂量依赖性。IWR-1可引起β-catenin和c-myc mRNA表达下降,β-catenin、c-myc蛋白表达下降,而Axin 1和p-β-catenin蛋白表达上升。结论 IWR-1可以抑制人肝癌细胞株Hep3B的增殖,其机制可能是通过抑制Wnt/β-catenin通路来实现。     

Ectopic Expression of miR-147 Inhibits Stem Cell Marker and Epithelial–Mesenchymal Transition (EMT)-Related Protein Expression in Colon Cancer Cells

Colon cancer is one of the most common cancers in the world. Epithelial-to-mesenchymal transition (EMT) is a crucial step in tumor progression and is also involved in the acquisition of stem cell-like properties. Some miRNAs have been shown to function as either tumor suppressors or oncogenes in colon cancer. Here we investigated the role of miR-147 in the regulation of the stem cell-like traits of colon cancer cells. We observed that miR-147 was downregulated in several colon cancer cell lines, and overexpressed miR-147 decreased the expression of cancer stem cell (CSC) markers OCT4, SOX2, and NANOG in the colon cancer cell lines HCT116 and SW480. Overexpressed miR-147 inhibited EMT by increasing the expression of epithelial markers E-cadherin and -catenin while decreasing the expression of mesenchymal markers fibronectin and vimentin. Moreover, activation of EMT by TGF- 1 treatment significantly counteracted the inhibitive effect of miR-147 on the expression of CSC markers OCT4, SOX2, and NANOG, supporting the idea that overexpressing miR-147 inhibited stem cell-like traits by suppressing EMT in colon cancer. In addition, we found that overexpressed miR-147 downregulated the expression of -catenin, c-myc, and survivin, which were related to the Wnt/ -catenin pathway. Moreover, treatment of miR-147 mimic-transfected cells with the Wnt/ -catenin pathway activator LiCl attenuated the inhibitive effect of the miR-147 mimic on the EMT and stem cell-like traits of colon cancer cells, indicating that ectopic expression of miR-147 inhibited stem cell-like traits in colon cancer cells by suppressing EMT via the Wnt/ -catenin pathway. In summary, our present study highlighted the crucial role of miR-147 in the inhibition of the stem cell-like traits of colon cancer cells and indicated that miR-147 could be a promising therapeutic target for colon cancer treatment.     

Profilin 2 Promotes Proliferation and Metastasis of Head and Neck Cancer Cells by Regulating PI3K/AKT/b-Catenin Signaling Pathway

Profilin 2 (PFN2) was found to be mainly expressed in neurons and involved in the development of the brain. In recent years, emerging evidence indicated that PFN2 is also significantly upregulated in various cancers including head and neck cancer (HNSC) and influences cancer cell proliferation, migration, and invasion. However, the role of PFN2 in HNSC development and progression remains unclear. The aim of our study was to investigate the role of PFN2 in the development of HNSC and its possible molecular mechanisms. Bioinformatics showed that increased expression of PFN2 in tumors correlated highly with poor prognosis of HNSC patients. Our results indicated that PFN2 was highly expressed in HNSC tissues and in HNSC cell lines. Knockdown of PFN2 inhibited proliferation, invasion, and migration of HNSC cells, while PFN2 overexpression produced the opposite effects. Using a nude mouse xenograft model, we substantiated the tumor-promoting effect of PFN2 on HNSC in vivo. Furthermore, we found that PFN2 downregulation reduced the phosphorylation of Akt and GSK-3 and reduced the expression of -catenin in HNSC cells. The opposite was observed when PFN2 was overexpressed. Collectively, these results suggest that PFN2 promotes the proliferation and metastasis of HNSC by activating the PI3K/Akt/ -catenin signaling pathway. Although further validation is needed, we speculate that PFN2 plays a crucial role in HNSC and may be a promising therapeutic target and prognostic biomarker.     

[ARTICLE WITHDRAWN] miR-630 Inhibits Epithelial-to-Mesenchymal Transition (EMT) by Regulating the Wnt/β-Catenin Pathway in Gastric Cancer Cells

THIS ARTICLE WAS WITHDRAWN BY THE PUBLISHERS IN OCTOBER 2020.     

香烟烟雾激活肺癌细胞Wnt/β-catenin信号途径的机制研究

背景与目的：香烟烟雾诱导的炎性反应可显著促进肺肿瘤的生长,本研究通过体内外实验明确香烟烟雾激活肺癌细胞Wnt/β-catenin信号途径的机制。方法：利用尾静脉注射鼠Lewis肺癌细胞方法建立小鼠肺癌模型,然后采用香烟烟雾对其进行刺激,诱发炎性反应。免疫组化方法检测小鼠和人肺肿瘤组织中CD68、肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)和无磷酸化β-catenin的表达。采用Transwell^®插入式细胞培养皿共培养人巨噬细胞和人肺癌细胞株A549细胞。采用蛋白质印迹法(Western blot)检测A549细胞中无磷酸化β-catenin的表达及磷酸化的GSK-3β和Akt蛋白的表达。结果：免疫组化结果显示,香烟烟雾刺激肺肿瘤负荷小鼠后,其肿瘤组织中巨噬细胞CD68⁺、TNF-α和无磷酸化β-catenin的表达均明显高于对照组。肺癌患者肿瘤组织中,CD68和TNF-α的阳性表达均明显高于正常组织。肺腺癌和鳞癌的无磷酸化β-catenin的染色阳性率分别为68.9%和62.2%,其阳性率与患者吸烟与否及肿瘤分期有关,差异有统计学意义(P均＜0.05),并且CD68⁺的细胞数在无磷酸化β-catenin染色阳性的标本中显著升高(P＜0.01)。Western blot检测结果显示,不同浓度香烟抽提物(cigarette smoke extract,CSE)刺激共培养细胞后,A549细胞中无磷酸化β-catenin的表达随着CSE浓度的增加而显著上升。在加入TNF-α中和抗体后,A549细胞中无磷酸化β-catenin的表达被抑制。用TNF-α处理A549细胞后,磷酸化的GSK-3β和Akt蛋白在2 h后表达增加,4 h后无磷酸化β-catenin蛋白表达也开始上调。结论：香烟烟雾诱导肺肿瘤组织中巨噬细胞释放炎性反应因子TNF-α,进而通过Akt/GSK-3β途径激活肿瘤细胞中Wnt/β-catenin信号通路。     

20(S)‐ginsenoside‐Rg3 reverses temozolomide resistance and restrains epithelial‐mesenchymal transition progression in glioblastoma

Glioblastoma multiforme (GBM) is one of the most malignant human intracranial tumors. Temozolomide (TMZ) is the primary alkylating agent for GBM patients. However, many GBM patients are resistant to TMZ. Therefore, patients with GBM urgently need more effective therapeutic options. 20(S)‐ginsenoside‐Rg3 (20(S)‐Rg3) is a natural chemical with anti‐tumor effects, but at present there is little understanding of its functional mechanism. Several research reports have demonstrated that O⁶‐methylguanine DNA‐methyltransferase (MGMT) repairs damaged DNA and contributes to TMZ resistance in gliomas. In addition, recent studies have shown that MGMT gene expression could be regulated by the Wnt/β‐catenin pathway. However, whether 20(S)‐Rg3 inhibits MGMT expression and augments chemosensitivity to Temozolomide (TMZ) in glioma cells remains unclear. In this study, we explored the modulating effects of 20(S)‐Rg3 on MGMT. We used glioma cell lines, primary cell strain (including T98G, U118 and GBM‐XX; all of them are MGMT‐positive glioma cell lines) and xenograft glioma models to examine whether 20(S)‐Rg3 increased the sensitivity to TMZ and to reveal the underlying mechanisms. We found that the MGMT expression was effectively downregulated by 20(S)‐Rg3 via the Wnt/β‐catenin pathway in glioma cell lines, and TMZ resistance was significantly reversed by 20(S)‐Rg3. Meanwhile, 20(S)‐Rg3 shows no obvious cytotoxicity at its effective dose and is well tolerated in vivo. In addition, we found that 20(S)‐Rg3 significantly restrains the epithelial‐mesenchymal transition (EMT) progression of glioma cells. Taken together, these results indicate that 20(S)‐Rg3 may be a novel agent to use in treatment of GBM, especially in TMZ‐resistant GBM with high MGMT expression.     

Prognostic relevance of epidermal growth factor receptor (EGF-R) and c-neu/erbB2 expression in glioblastomas (GBMs)

Summary Seventeen untreated primary adult glioblastomas were analyzed using immunocytochemistry for the expression of EGF-R, c-neu/erbB2, TGF-, and phosphotyrosine. Patients were divided by median survival into long-term or short-term survivors (LTS, N=10, median > 4 years; versus STS, N=7, median 61 weeks). There were no significant differences between the two groups in terms of age, extent of resection, post-operative Karnofsky status, or treatment. Diagnostic sections from each tumor were stained with antibodies to EGF-R, c-neu/erbB2, TGF- and phosphotyrosine. Double-labelling for TGF- and EGF-R was also performed. All 10/10 LTS were considered to be EGF-R negative/scant, while 4/7 STS were EGF-R positive. EGF-R negativity significantly correlated with long-term survival. The differences in c-neu/erbB2 expression did not reach significance. However, 4/7 STS were positive for both proteins and 76% of the 17 cases were either double negative or positive for EGF-R and c-neu/erbB2. TGF- and phosphotyrosine were frequently expressed, but neither were prognostic. Recurrent tumors were studied in 7 STS. EGF-R expression was increased in 4/7 of these cases and c-neu/erbB2 was increased in all 7 cases, compared to the pretreatment baselines. Increased expression of these proteins in glioblastomas may be associated with aggressive clinical behavior and treatment resistance.     

RNA激活上调p21~(WAF1/CIP1)基因表达对肺癌细胞增殖和凋亡影响的研究

目的:探讨应用RNA激活(RNAa)技术上调p21WAF1/CIP1(p21)表达对人肺癌H441细胞增殖和凋亡的影响。方法:设计靶向抑癌基因p21WAF1/CIP1启动子DNA序列互补的双链RNA分子(dsp21),转染H441细胞后应用半定量逆转录聚合酶链反应(RT-PCR)及蛋白质印迹法检测p21WAF1/CIP1基因mRNA及蛋白质的表达变化,噻唑蓝(MTT)法检测H441细胞增殖速度的变化,流式细胞仪检测H441细胞凋亡率的变化。结果:dsp21转染H441细胞72 h后p21WAF1/CIP1表达显著上调。RT-PCR灰度比值结果显示,空白对照组、阴性对照组和实验组p21WAF1/CIP1mRNA相对表达量分别为(38.1±2.5)%、(33.5±3.9)%和(96.5±2.3)%,差异有统计学意义,F=87.0,P<0.01;蛋白质印迹法结果显示,空白对照组、阴性对照组和实验组p21WAF1/CIP1蛋白相对表达量分别为(45.7±2.2)%、(43.2±3.1)%和(93.6±2.5)%,差异有统计学意义,F=79.0,P<0.01;经dsp21作用的H441细胞增殖受到明显抑制,凋亡率较对照...