MicroRNA-335 inhibits invasion and metastasis of colorectal cancer by targeting ZEB2

To determine whether chemotherapy treatment at least 6 months prior to the detection of hepatic steatosis is associated with advanced hepatic fibrosis. Demographics, comorbid conditions, and laboratory data for cancer patients with hepatic steatosis were reviewed. The primary end point of this study was a low probability of fibrosis as calculated by the AST-to-platelet ratio index (APRI)—a surrogate for the absence of histologic bridging fibrosis and/or cirrhosis. Of 279 patients, 117 (41.9 %) were treated with chemotherapy and 197 (66.3 %) had a low probability of fibrosis by APRI. A smaller proportion of patients treated with chemotherapy had a low probability of hepatic fibrosis compared with untreated patients (64.1 vs. 75.3 %, p = 0.04). On multivariable analysis, chemotherapy treatment was a negative predictive factor for a low probability of fibrosis (OR 0.366 [95 % CI 0.184–0.708], p < 0.01). Among chemotherapy-treated patients, 75 (64.1 %) had a low probability of fibrosis. There were no differences in chemotherapy duration (mean 7.8 vs. 7.5 cycles) and interval from last dose to steatosis diagnosis (24.3 vs. 21.4 months) between patients with and without a low probability of fibrosis. A smaller proportion of patients treated with irinotecan or 5-fluorouracil had a low probability of fibrosis (37.3 vs. 66.7 %, p = 0.04). On multivariable analysis, irinotecan or 5-fluorouracil treatment was a negative predictive factor for low probability of fibrosis (OR 0.277 [95 % CI 0.091–0.779], p = 0.02). Prior chemotherapy treatment, especially with 5-fluorouracil or irinotecan, is a negative predictor for the absence of advanced hepatic fibrosis among patients with steatosis.     

MicroRNA-9 up-regulation is involved in colorectal cancer metastasis via promoting cell motility

Human microRNA-9 (miR-9) has been reported to be involved in the metastasis of several malignancies including brain breast cancer. However, its role in the metastasis of colorectal cancer (CRC) remains to be revealed. Here, we evaluated miR-9 expression in metastatic CRC and investigated its effects on the motility and proliferation of RKO cells. The expressions of miR-9 in 15 primary CRC specimens without distant metastasis (NM group) and 10 primary CRC specimens (M group) with distant metastasis (M group) were determined by quantitative real-time PCR. The alternations in the motility and morphology of RKO cells before and after miR-9 transfection were analyzed by migration assay and F-actin staining. The relationship between miR-9 and α-catenin was identified by Western blotting. Cell growth was examined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide) assay. Significant difference of miR-9 expression was observed in M group compared to the NM group (P < 0.001). Ectopic expression of miR-9 enhanced the motility of RKO cells as well as changed their morphological appearance, while cell growth remained unchanged. The overexpression of miR-9 could also down-regulate α-catenin expression. These data suggest that miR-9 may potentially participate in the metastatic process of CRC though facilitating cell motility.     

MicroRNA-145 inhibits tumour growth and metastasis in colorectal cancer by targeting fascin-1

Background:

Recent studies have reported miR-145 dysregulated in colorectal cancer (CRC). In this study, miR-145 profiles were compared between CRC and corresponding non-tumour tissues.

Methods:

The expression levels of miR-145 were analysed in CRC cell lines and tumour tissues by real-time PCR. A luciferase reporter assay confirmed direct targets. The functional effects of miR-145 were examined in transfected CRC cells in vitro and in vivo using established assays.

Results:

Downregulation of miR-145 was detected in most primary CRC tumours, and was significantly correlated with a more aggressive phenotype of CRC in patients. In CRC cell lines, ectopic overexpression of miR-145 inhibited cell proliferation, motility and invasion in vitro. Stable overexpression of miR-145 suppressed tumour growth and pulmonary metastasis in vivo. Further studies indicated that miR-145 may directly interact with the 3′-untranslated region (3′-UTR) of Fascin-1 messenger RNA (mRNA), downregulating its mRNA and protein expression levels. In clinical specimens, Fascin-1 expression was negatively correlated with miR-145 expression.

Conclusions:

MiR-145 has a critical role in the inhibition of invasive and metastatic capacities of CRC, probably through directly targeting Fascin-1. This miRNA may be involved in the development and progression of CRC.     

Beta-hydroxyisovalerylshikonin and cisplatin act synergistically to inhibit growth and to induce apoptosis of human lung cancer DMS114 cells via a tyrosine kinase-dependent pathway

beta-Hydroxyisovalerylshikonin (beta-HIVS) and cisplatin (CDDP) had a synergistic growth-inhibitory effect on cultured human small-cell lung carcinoma DMS114 cells, as well as on human leukemia U937 and epidermoid carcinoma A431 cells, while beta-HIVS and CDDP alone at the same respective concentrations had little effect. Growth inhibition was accompanied by induction of apoptosis, as determined by an ELISA for the detection of cell death and the TUNEL assay. Using phosphotyrosine-specific antibodies (PY20), we observed that tyrosine kinase activity in DMS114 cells was inhibited by treatment with beta-HIVS and CDDP together. The tyrosine kinase activity of isolated Src and that of isolated receptors for epidermal growth factor were also inhibited by the two agents together. The synergistic effects of the growth of DMS114 cells of beta-HIVS and CDDP were not due simply to the intracellular accumulation of CDDP or to levels of DNA adducts. Our data suggest that the synergistic effect on the growth of DMS114 cells of beta-HIVS and CDDP might be a result of the inhibition of a tyrosine kinase-dependent pathway.     

1Alpha,25dihydroxyvitamin D3 and platinum drugs act synergistically to inhibit the growth of prostate cancer cell lines.

The majority of men who die from prostate cancer (PC) have hormone-refractory disease. To date, chemotherapeutic agents have had little or no impact on the survival of such patients. To explore a new approach for the treatment of hormone-refractory PC, we examined the combination effects of cis- or carboplatin with vitamin D. 1alpha,25-Dihydroxyvitamin D3 (1alpha,25(OH)2D3) and its synthetic analogue, Ro 25-6760, have an antiproliferative effect on some prostate cancer cell lines. Consequently, the growth-inhibitory effects of the drugs were measured, both singularly and in combination with cis- or carboplatin, on PC cells. Our results show that although each of the drugs alone displayed antiproliferative activity, the growth inhibition of PC cells was further enhanced by the combination of 1alpha,25(OH)2D3 or Ro 25-6760 and either platinum agent. The greatest enhancement of inhibition occurred using smaller concentrations of the platinum compound in combination with higher concentrations of 1alpha,25(OH)2D3. Isobologram analysis revealed that 1alpha,25(OH)2D3 and platinum acted in a synergistic manner to inhibit the growth of PC cells. Our findings suggest that there is potential clinical value in combining 1alpha,25(OH)2D3 with platinum compounds for the treatment of advanced-stage human PC.     

Oncolytic adenovirus mediated Survivin RNA interference and 5-fluorouracil synergistically suppress the lymphatic metastasis of colorectal cancer

Colorectal cancer is one of the leading malignancies in the world. The mortality is mainly caused by tumor metastasis. It is urgent to find new therapeutics against metastatic colorectal cancer. We constructed an adenovirus carrying a Survivin targeted shRNA, tested its effects alone or with 5-fluorouracil (5-FU) both in vitro and in a nude mouse xenograft model. Results showed that the recombinant adenovirus reduced the expression of Survivin effectively, and administration of virus together with 5-FU inhibited cancer cell metastasis both in vitro and in vivo at concentrations at which each agent alone was ineffective. We conclude Survivin targeted virotherapy together with 5-FU chemotherapy may be a promising treatment for metastatic colorectal cancer.     

Metformin and temozolomide act synergistically to inhibit growth of glioma cells and glioma stem cells in vitro and in vivo

Glioblastoma (GBM) is the most frequent and aggressive brain tumor in adults. In spite of advances in diagnosis and therapy, the prognosis of patients with GBM has remained dismal. The fast recurrence and multi-drug resistance are some of the key challenges in combating brain tumors. Glioma stem cells (GSCs) which are considered the source of relapse and chemoresistance, the need for more effective therapeutic options is overwhelming. In our present work, we found that combined treatment with temozolomide (TMZ) and metformin (MET) synergistically inhibited proliferation and induced apoptosis in both glioma cells and GSCs. Combination of TMZ and MET significantly reduced the secondary gliosphere formation and expansion of GSCs. We first demonstrated that MET effectively inhibited the AKT activation induced by TMZ, and a combination of both drugs led to enhanced reduction of mTOR, 4EBP1 and S6K phosphorylation. In addition, the combination of the two drugs was accompanied with a powerful AMP-activated protein kinase (AMPK) activation, while this pathway is not determinant. Xenografts performed in nude mice demonstrate in vivo demonstrated that combined treatment significantly reduced tumor growth rates and prolonged median survival of tumor-bearing mice. In conclusion, TMZ in combination with MET synergistically inhibits the GSCs proliferation through downregulation of AKT-mTOR signaling pathway. The combined treatment of two drugs inhibits GSCs self-renewal capability and partly eliminates GSCs in vitro and in vivo. This combined treatment could be a promising option for patients with advanced GBM.     

蟾毒灵与顺铂协同抑制乳腺癌MCF-7细胞增殖的机制探讨

目的 探讨蟾毒灵（Bufalin）联合顺铂对乳腺癌MCF-7细胞增殖及凋亡的影响,并探讨可能的协同机制。方法 采用MTT法检测空白对照组（仅培养液）、单纯细胞对照组和实验组的细胞增殖率,实验组加入不同浓度的顺铂或Bufalin单药或以固定比例（Bufalin∶顺铂=1 nmol／L∶1 μmol／L）联合作用24 h。分别采用20 nmol／L Bufalin、20 μmol／L顺铂单药或联合作用24 h,用流式细胞术检测细胞凋亡,并用Western Blotting检测活化的Met（p-Met）、Met、活化的Src（p-Src）、Src、PARP及其裂解cleaved PARP蛋白的表达。结果 MTT法检测显示,顺铂和Bufalin均以剂量依赖方式抑制乳腺癌MCF-7细胞增殖。在顺铂≥0.1 μmol／L和Bufalin≥0.1 nmol／L时两药联合作用可协同抑制MCF-7细胞增殖（0＜CI＜0.433）。流式细胞术检测显示,20 μmol／L顺铂作用24 h可诱导（10.7±4.8）％ 的MCF-7细胞凋亡,而20 nmol／L Bufalin作用24 h未明显诱导细胞凋亡,20 nmol／L Bufalin与20 μmol／L顺铂联合作用24 h后可诱导（40.8±8.5）％的MCF-7细胞凋亡。Western Blotting检测显示,顺铂作用后可导致Met和Src活化,Bufalin与顺铂联合作用后可抑制顺铂诱导的Met和Src活化,同时上调PARP裂解。结论 Bufalin可能通过抑制顺铂诱导的Met和Src活化与顺铂协同抑制MCF-7细胞增殖,增强顺铂诱导的细胞凋亡。     

MicroRNA-29a promotes colorectal cancer metastasis by regulating matrix metalloproteinase 2 and E-cadherin via KLF4

Background:

Growing evidence suggests that miR-29a has an important role in regulating tumourigenesis and development of various types of cancer. However, the role and the underlying mechanism of miR-29a in colorectal cancer (CRC) remain largely unknown.

Methods:

MiR-29a targeted gene was identified by the luciferase assay and western blot. MiR-29a function was analysed by invasion assays and the orthotopic transplantation mouse model. The miR-29a pathway was assayed by real-time PCR, western blot and chip analysis.

Results:

KLF4 was identified as a direct target gene of miR-29a. MiR-29a promoted CRC cell invasion, which was blocked by re-expression of KLF4. In addition, MMP2 was identified as a novel direct target of KLF4. Both miR-29a overexpression and KLF4 knockdown promoted MMP2 expression but inhibited E-cadherin expression. Furthermore, clinical data indicated that both miR-29a high expression and KLF4 mRNA low expression were associated with metastasis and poor prognosis in CRC patients, and KLF4 protein expression was inversely correlated with MMP2 but positively correlated with E-cad protein expression.

Conclusion:

Increased expression of miR-29a promoted CRC metastasis by regulating MMP2/E-cad through direct targeting KLF4, which highlights the potential of the miR-29a inhibitor as a novel agent against CRC metastasis.     

Urokinase receptor and vascular endothelial growth factor are synergistically associated with the liver metastasis of colorectal cancer.

Considering recent findings that the urokinase plasminogen activation (PA) system is involved in invasion and vascular endothelial growth factor (VEGF) is involved in angiogenesis of colorectal cancer, we evaluated these factors in the liver metastasis of primary colorectal cancer. Cancer tissues from 71 colorectal cancer patients were assayed quantitatively for antigen levels of urokinase type plasminogen activator (uPA), uPA receptor (uPAR), and plasminogen activator inhibitor-1 and -2 (PAI-1, PAI-2), and were also assayed immunohistochemically for expression of VEGF protein. Among the PA system factors, both the levels of uPAR and PAI-1 were significantly higher in larger tumors than in smaller ones, and were also significantly higher in tumors that invaded subserosa, serosa or adjacent organs than in mucosal, submucosal tumors or in tumors that invaded the muscle layer. The uPAR levels were significantly higher in tumors with liver metastasis than in those without. VEGF overexpression was significantly more frequent in tumors with lymph node involvement or liver metastasis than in those without. Among the PA system factors, the uPAR levels were significantly higher in tumors with VEGF overexpression and a multivariate analysis revealed that high uPA level and VEGF overexpression were independent risk factors for liver metastasis. The combination of high uPAR level and overexpression of VEGF was associated with the worst prognosis in patients with colorectal cancer. These results suggest that uPAR and VEGF might contribute synergistically to the liver metastasis of colorectal cancer.     

Microbiota in colorectal cancer related to liver metastasis

The prevalence of colorectal cancer(CRC) is increasing annually and metastasis is the principal cause of death in patients with CRC, with the liver being the most frequently affected site. Many studies have shown a strong interplay between the gut flora, particularly Fusobacterium nucleatum(F. nucleatum), Escherichia coli, and Bacteroides fragilis, and the development of gut tumors. Some strains can induce gut inflammation and produce toxins that directly harm gut epithelial cells, ultimately ac...     

CXCR4抑制剂AMD3100对结肠癌肝转移的实验研究

目的:研究CXCR4特异性抑制剂AMD3100治疗结肠癌肝转移的效果.方法:建立结肠癌肝转移动物模型.将18只模型裸鼠随机分为对照组(0.1 mL生理盐水)、低剂量治疗组(AMD3100,4 mg/kg)和高剂量治疗组(AMD3100,6 mg/kg),每组6只,治疗6周后评价疗效.结果:对照组、低剂量治疗组和高剂量治疗组脾脏原发瘤直径大小分别为(13.83±7.73)、(5.17±0.75)和(2.83±1.33) mm(治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).对照组、低剂量治疗组和高剂量治疗组肝脏转移瘤直径大小分别为 (3.42±2.24)、(2.26±1.12)和(1.62±0.88) mm(治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).对照组、低剂量治疗组和高剂量治疗组肝脏表面转移结节计数分别为21.00±3.90、13.00±1.79和2.17±1.37 (治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).结论:CXCR4特异性抑制剂AMD3100可以降低模型动物的结肠癌肝转移程度,抑制肝脏转移瘤的生长,且呈浓度依赖性;SDF-1/CXCR4轴在结肠癌肝脏转移中具有重要作用.     

Unusual metastasis in colorectal cancer

Metastasis from colorectal carcinoma occurs by either lymphatic or hematogenous spread. The most common sites of colorectal metastasis are the liver and lung. Involvement of the skin, muscles and bones are quite rare. The prognosis in such patients is usually poor. Herewith, we are reporting a case of colonic carcinoma who had cutaneous metastasis, muscular involvement and diffuse skeletal metastasis. At the end, she had brain metastasis, but liver and lung involvement was not observed till the end.     

MicroRNA-17-5p promotes chemotherapeutic drug resistance and tumour metastasis of colorectal cancer by repressing PTEN expression

Background: Colorectal cancer (CRC) is one of the most common cancers worldwide, especially in Western countries. Although chemotherapy is used as an adjuvant or as a palliative treatment, drug resistance poses a great challenge. This study intended to identify biomarkers as predictive factors for chemotherapy.Patients and methods: By microarray analysis, we studied miRNAs expression profiles in CRC patient, comparing chemoresistant and chemosensitive groups. The miRNAs of interest were validated and the impact on clinical outcomes was assessed in a cohort of 295 patients. To search for potential targets of these miRNAs, tissue samples were subject to in situ hybridization and immunohistochemistry analysis. Colorectal adenocarcinoma cells were also used for in vitro experimentation, where cellular invasiveness and drug resistance were examined in miRNA-transfected cells.Results: The expression level of miRNA-17-5p was found increased in chemoresistant patients. Significantly higher expression levels of miR-17-5p were found in CRC patients with distant metastases and higher clinical stages. Kaplan-Meier analysis showed that CRC patients with higher levels of miR-17-5p had reduced survival, especially in patients who had previously received chemotherapy. Overexpression of miR-17-5p promoted COLO205 cell invasiveness. We found that PTEN was a target of miR-17-5p in the colon cancer cells, and their context-specific interactions were responsible for multiple drug-resistance. Chemotherapy was found to increase the expression levels of miR-17-5p, which further repressed PTEN levels, contributing to the development of chemo-resistance.Conclusions: MiR-17-5p is a predictive factor for chemotherapy response and a prognostic factor for overall survival in CRC, which is due to its regulation of PTEN expression.     

MicroRNA-183抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移

目的:探讨microRNA183 （miRNA-183）对结直肠癌肿瘤细胞增殖、侵袭及迁移的影响。方法:使用慢病毒技术,分别转染HT29细胞株,建立稳定过表达细胞株miRNA-183 mimics,抑制miRNA-183表达的稳定细胞株miRNA-183 inhibitor以及阴性对照组negative control。qRT-PCR检测各组细胞miRNA-183的表达,Western blot检测miRNA-183下游靶基因PDCD4蛋白水平表达变化,CCK8增殖实验检测各组细胞的增殖情况,同时以划痕实验和Transwell实验分析转染miRNA-183 mimics和miRNA-183 inhibitor对HT29细胞迁移、侵袭能力的影响。结果:与阴性对照组相比,miRNA-183 mimics组细胞增殖速度明显下降,其迁移和侵袭能力也显著降低,差异均具有统计学意义（P<0.05）;miRNA-183下游靶基因PDCD4蛋白水平表达明显增加。结论:miRNA-183可有效地抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移。     

MicroRNA-497 Suppresses Proliferation and Induces Apoptosis in Prostate Cancer Cells

MicroRNAs (miRNAs) are a class of endogenously expressed small, non-coding, single-stranded RNAsthat negatively regulate gene expression, mainly by binding to 3’- untranslated regions (3’UTR) of their targetmessenger RNAs (mRNAs), which cause blocks of translation and/or mRNA cleavage. Recently, miRNAprofilingstudies demonstrated the microRNA-497 (miR-497) level to be down-regulated in all prostate carcinomascompared with BPH samples. The purpose of this study was to investigate the potential role of miR-497 inhuman prostate cancer. Proliferation, cell cycle and apoptosis assays were conducted to explore the potentialfunction of miR-497 in human prostate cancer cells. Results showed that miR-497 suppressed cellular growth andinitiated G0/G1 phase arrest of LNCaP and PC-3 cells. We also observed that miR-497 increased the percentageof apoptotic cells by increasing caspase-3/7 activity. Taken together, our results demonstrated that miR-497 caninhibit growth and induce apoptosis by caspase-3 activation in prostate cancer cells, which suggest its use as apotential therapeutic target in the future.     

American ginseng and breast cancer therapeutic agents synergistically inhibit MCF-7 breast cancer cell growth

BACKGROUND AND OBJECTIVES: American ginseng (Panax quinquefolius L.) purportedly alleviates menopause symptoms because of putative estrogenicity. METHODS: Using a standardized American ginseng (AG) extract in MCF-7 breast cancer cells, the objectives were to evaluate the ability of AG to induce the estrogen- regulated gene pS2 by Northern blot analysis, determine the effect on cell growth using the MTT assay, and evaluate the cell cycle effects by flow cytometry. RESULTS: AG and estradiol equivalently induced RNA expression of pS2. AG, in contrast to estradiol, caused a dose-dependent decrease in cell proliferation (P < 0.005). AG had no adverse effect on the cell cycle while estradiol significantly increased the proliferative phase (percent S-phase) and decreased the resting phase (G(0)-G(1) phase) (P < 0.005). Concurrent use of AG and breast cancer therapeutic agents resulted in a significant (P < 0.005) suppression of cell growth for most drugs evaluated. CONCLUSIONS: In vitro use of AG and breast cancer therapeutics synergistically inhibited cancer cell growth.