HCC-MSCs significantly induced the EMT pathway of HCC cells.

RNA pulldown assays followed by mass spectrum indicated that lncRNA-MUF can bind with ANXA2.

RNA pulldown assays followed by mass spectrum indicated that lncRNA-MUF can bind with ANXA2.

Human lncRNA-MUF sequence has no coding potentiality.

ANXA2 modulates beta-catenin-target genes and can interact with GSK3beta.

LncRNA-MUF acts as a ceRNA to modulate the function of miR-34a.

HCC-MSCs induced EMT and upregulated lncRNA-MUF,ANXA2 interacted with GSK3 beta,lncRNA-MUF acted as a ceRNA of miR34a and other detailed methods

LncRNA-MUF is upregulated in HCC cells by HCC-MSCs

Evasion of growth suppression is among the prominent hallmarks of cancer. Phosphatase and tensin homolog (PTEN) and p53 tumor-suppressive pathways are compromised in most human cancers, including glioblastoma (GB). Hence, these signaling pathways are an ideal point of focus for novel cancer therapeutics. Recombinant viruses can selectivity kill cancer cells and carry therapeutic genes to tumors. Specifically, oncolytic viruses (OV) have been successfully employed for gene delivery in GB animal models and showed potential to neutralize immunosuppression at the tumor site. However, the associated systemic immunogenicity, inefficient transduction of GB cells, and inadequate distribution to metastatic tumors have been the major bottlenecks in clinical studies. Mesenchymal stem cells (MSCs), with tumor-tropic properties and immune privilege, can improve OVs targeting. Remarkably, combining the two approaches can address their individual issues. Herein, we summarize findings to advocate the reactivation of tumor suppressors p53 and PTEN in GB treatment and use MSCs as a “Trojan horse” to carry oncolytic viral cargo to disseminated tumor beds. The integration of MSCs and OVs can emerge as the new paradigm in cancer treatment. Graphical abstract

Objective To investigate the status quo of illness perception and quality of life in thyroid cancer patients, analyze the factors affecting the quality of life. Methods A total of 210 cases of thyroid cancer patients were selected by convenience sampling. The Chinese version of the Brief Illness Perceptions Questionnaire (BIPQ), the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core30 (EORTC QLQ-C30) and Thyroid Cancer–Specific Quality of Life Questionnaire (THYCA-QoL) were used to collect data. Results The patients' illness perception score was (40. 27±8. 46), and the overall health status score was (55. 91±22. 67). Illness perception was negatively correlated with global health status (r=-0. 15, P＜0. 05). Multiple stepwise regression analysis showed that there are totally 9 independent factors affecting the quality of life, including age, gender, educational level, average monthly income, treatment methods, emotions representation and thyroid cancer–specific symptom such as felt chilly, tingling hands, sympathetic (P＜0. 05). Conclusion Thyroid cancer patients have moderate illness perception, the higher the level of illness perception, the worse the quality of life. Age, gender, level of education, average monthly income, treatment methods, emotions representation and thyroid cancer–specific symptom such as felt chilly, tingling hands, sympathetic are influential factors towards quality of life.

Objective To assess the awareness, attitude and barriers of colorectal cancer screening among high-risk populations in China. Design A cross-sectional study was employed. Setting This study was conducted in nine hospitals in Hunan province, China. Participants Individuals with a high-risk for colorectal cancer were interviewed using a pretested structured questionnaire. Primary and secondary outcome measures Knowledge, attitude towards colorectal cancer screening, sociodemographic factors associated with screening knowledge and behaviour and barriers of colorectal cancer screening. Results This study included 684 participants. The mean knowledge score was 11.86/24 (SD 4.84). But over 70% of them held a positive attitude towards screening. Only 13.3% had undergone colorectal cancer screening. Independent factors related to knowledge were education level of college or above, working as a white collar, higher income, having health insurance, having seen a doctor in the past year and with a high perceived risk (p<0.05). Factors independently associated with screening behaviour included personal history of colorectal disease, having seen a doctor in the past year, previous discussion of colorectal cancer screening, high perceived risk and better knowledge (p<0.05). Main reasons for not undergoing screening were no symptoms or discomfort (71.1%), never having thought of the disease or screening (67.4%) and no doctor advised me (29.8%). Conclusion In China, the majority of high-risk people had deficient knowledge and had never undergone colorectal cancer screening. But most of them held a positive attitude towards the benefits of colorectal cancer screening. This has promising implications to design targeted educational campaigns and establish screening programmes to improve colorectal cancer awareness and screening participation. Healthcare professionals should advise high-risk individuals to participate in screening and inform them about cancer risk.

Purpose: The study aims to lay a foundational probe for the thorough application microfluidic chips in brain function research with microfluidic chips. Neuron slide culture is a common culture method in vitro, and the microfluidic chip with the artificial network pattern not only can realize neuron cells 3 D culture in vitro, but also limit the extension space of neurite outgrow. Materials and Methods: In order to analyze the differences of hippocampal cells neurite growth length between the 3 D chips and the common 2 D culture, the experiments utilized statistical analysis method analyzing the length of the hippocampus neuron neurite of 3 days, 5 days and 7 days, respectively, with the common glass slide 2 D culture method and the microfluidic chip 3 D culture in vitro. Results: The results showed that there was no significant difference in the neurite length after 3 days. However, there was a significant difference after 5 days and 7 days. It can be seen that the microfluidic chip with artificial network pattern has limitations to the growth of neurite after 5 days. Conclusions: We concluded that the growth state of hippocampal cells in the restricted 3 D space is different from that of conventional 2 D culture.It showed that the artificial network pattern design has limited the growth space of the dendrites but also affected its growth.

腺病毒(Adenovirus,AdV)是常见的基因治疗载体,已有的腺病毒下游纯化工艺通常采取阴离子交换层析搭配分子筛的方式,该方法存在耗时长、成本高、难以放大并且所获得的腺病毒中具有感染性的腺病毒比例较低的问题,本研究旨在探究并优化一步层析法,纯化得到高滴度的5型腺病毒.首先在5L生物反应器中利用HEK-293.2sus细胞扩增5型腺病毒,通过对细胞沉淀的反复冻融释放病毒,利用过滤和超滤技术去除部分杂质,并通过添加核酸酶降低料液的黏稠度,最后利用新型复合填料Capto core 700或Fractogel填料进行纯化.纯化后的病毒利用50％组织细胞感染量(Median tissue culture infective dose,TCID50)测定方法测定病毒滴度,利用PCR-荧光探针法检测宿主DNA的残留以及酶联免疫法检测宿主蛋白的残留以判断除杂效果.两种一步层析法纯化腺病毒的回收率均高于传统的两步法,但在宿主蛋白和宿主DNA的去除效果上Capto core700的结果均不如Fractogel填料与超滤置换的结合,所以利用超滤和Fractogel结合的纯化方式更适合获得临床级别的腺病毒.

Hypermethylation of gene promoter has been indicated for the contribution of gene silencing, and DNA demethylating drugs, such as 5-aza-2'-deoxycytidine (DAC), has been used clinically for cancer treatment. However, the reason why a proportion of genes with hypermethylated promoter exhibit high expression levels remains unclear and this drug is not much successful as expected in use. Furthermore, CpG islands (CGIs) are found to be located in not only promotors, but also in gene bodies. By RNA-seq and reduced representation bisulfite sequencing, we found the mismatch between the level of promoter methylation and gene expression. By chromatin Immunoprecipitation-quantitative polymerase chain reaction and luciferase reporter assay, we identified putative promoters in gene body, and proved the activities of putative promoters were affected by the methylation level of the CGI nearby. DAC can reverse the DNA hypermethylation at promoter CGIs effectively but not the CGIs in gene body. We also found that TET1 could demethylate CGIs both in promoter and gene body. Furthermore, we revealed a novel mechanism that H3K36me3 could affect the activity of putative promoter, and 5hmC recruited MeCP2 and CREB1 as a coactivator to SETD2 promoter, to enhance its gene expression and result in increased H3K36me3 in gene body. Our results concluded that putative promoters existed in the gene bodies, and TET1 could influence the transcriptional activity of putative promoters by intragenic demethylation.

Purpose Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and ranked top in terms of incidence and mortality in men and women. Recently, improvements in treatment approaches for NSCLC have reported, but still, there is a need to devise innovative treatment strategies, especially to manage the advanced and metastatic stage of NSCLC. Aloperine (ALO), an herbal alkaloid, has exerted anti-cancer effects in many cancers. However, the use of any chemotherapeutic agents is dose limited due to possible adverse effects and drug-resistance issues. Therefore, a combination of chemotherapy with viral-based targeted gene therapy may provide a novel treatment strategy for NSCLC. Methods/results In this study, the results of the MTT and flow cytometry-based assays showed that Aloperine-Adbic (adenoviral vector expressing p14(ARF)/p53) combined treatment on NSCLC cells synergistically produced anti-proliferative effects, induced apoptosis, and arrested cell cycle at the G1 phase. Furthermore, the expression analysis suggested that the p53/p21 pathway might contribute to achieving aforesaid cytotoxic effects. The ALO-Adbic combined treatment prolonged the percent survival of NSCLC xenograft models. Conclusion In conclusion, ALO-Adbic combination can produce synergistic anti-cancer effects at low doses, and may offer a more effective and less toxic new treatment strategy for NSCLC.

Cell fusion plays a crucial role in cancer progression and leads to massive aberrant changes in chromosome and gene expression involved in tumor metastasis. Cancer cells can fuse with many cell types, including stromal cells, epithelial cells, macrophages, and endothelial cells. Mesenchymal stem cells (MSCs) have been reported to migrate and incorporate into tumor sites during cancer progression. However, the underlying mechanism of stem cell fusion in tumor metastasis has not been fully deciphered. In this research, we established a cell fusion model between lung cancer cells and MSCs in vitro. We found that the hybrid cells showed enhanced metastatic capacity with increased expression of MMP-2 and MMP-9, whereas the proliferation ability was inhibited and cell cycle was blocked in the G0 /G1 phase with elevated expression of p21, p27, and p53. Moreover, the hybrid cells lost epithelial morphology and exhibited an epithelial-mesenchymal transition (EMT) change with downregulation of E-cadherin and upregulation of N-cadherin, Vimentin, α-SMA and Fibronectin1. Meanwhile, the expressions of EMT transcription factors, including Snail1, Slug, Twist1, Zeb1, and Zeb2, were also increased in hybrid cells. More important, the fusion hybrids acquired stem cell-like properties, which exhibited increased expression stem cell transcription factors Oct4, Sox2, Nanog, Kif4 as well as Bmi1. Taken together, our results suggested that cell fusion between lung cancer cells and MSCs offered enhanced metastatic capacity and characteristics of cancer stem cell by undergoing EMT. This study will contribute to explaning the origin of lung cancer stem cells and to elucidate the role of cell fusion in cancer metastasis.

BACKGROUND:There is an urgent need for targeted biological therapies for prostate cancer with greater efficacy and less toxicity, particularly for metastatic disease, where current therapies are not curative. Therapeutic adenoviral vectors or oncolytic adenoviruses offer the possibility of a competent, nontoxic therapeutic alternative for prostate cancer. However, free viral particles must be delivered locally, an approach that does not address metastatic disease, and they display poor tumor penetration. To fully exploit the potential of these vectors, we must develop methods that improve intratumoral dissemination and allow for systemic delivery. This study establishes a proof-of-principle rationale for a novel human mesenchymal stem (stromal) cell-based approach to improving vector delivery to tumors.METHODS/RESULTS:We have generated mesenchymal stem cell-derived packaging cells for adenoviruses (E1-modified mesenchymal stem cells) by modifying human mesenchymal stem cells with the adenovirus (type C) E1A/B genes needed for viral replication. Using cell-based assays, we have demonstrated that two adenoviral vectors, replication-defective adenovirus expressing p14 and p53 or conditionally replicating oncolytic adenovirus, packaged by E1A/B-modified mesenchymal stem cells, suppress the growth of prostate cancer cells in culture. Using subcutaneous xenograft models for human prostate cancer in mice, we have shown that E1A/B-modified mesenchymal stem cells display tumor tropism in tumor-bearing nude mice, that E1A/B-modified mesenchymal stem cells disseminate well within tumors, and that replication-defective adenovirus expressing p14 and p53 or conditionally replicating oncolytic adenovirus-loaded E1-modified mesenchymal stem cells suppresses tumor growth in mice.CONCLUSION:The results show that this approach, if optimized, could circumvent the obstacles to efficient gene delivery encountered with current gene delivery approaches and provide an effective, nontoxic therapeutic alternative for metastatic disease.

Cell fusion is a highly regulated process involved in cancer development, tissue regeneration and other physiological and pathological events. Many studies have shown that cancer cells can fuse with different types of cells such as mesenchymal stem cells (MSCs) and macrophages, which are behaved as two important fusogenic candidates in the tumor microenvironment. However, the underlying mechanisms of cell fusion between macrophages and malignant cells in cancer progression has not been fully clarified. The aim of the present study was to investigate the effects and mechanisms of cell fusion between macrophages and breast cancer cells on tumorigenesis and metastasis. Our results indicated that the hybrids exhibited enhanced proliferation, colony formation, migration and invasion capabilities, as well as suppressed apoptosis compared with parental breast cancer cells. Moreover, the hybrid cells displayed EMT with a significant downregulation of E-cadherin and upregulation of N-cadherin, Vimentin and Snail, as well as an obviously increased expression of MMP-2, MMP-9, uPA and S100A4. Mechanistically, we found that the TCF/LEF transcription factor activity of Wnt/β-catenin pathway and the expression of its downstream target genes including cyclin D1 and c-Myc were increased in the hybrid cells. Furthermore, our data confirmed that the promoting effects of fusion of macrophages on breast cancer cell proliferation, migration and invasion could be blocked by treatment with XAV-939, a Wnt/β-catenin signaling pathway inhibitor. In conclusion, our findings demonstrate that fusion of macrophages promotes proliferation, migration and invasion of breast cancer cells through activating EMT and Wnt/β-catenin signaling pathway. Our current study will further contribute to elucidate the mechanism of cell fusion in tumorigenesis and metastasis, and to develop a new therapeutic strategy for breast cancer treatment.

目的 研究姜黄素与5-FU联用对人结肠癌细胞自噬及对YAP的影响.方法 姜黄素、5-FU单独应用及两者联用作用于人结肠癌HCT116细胞和过表达YAP的结肠癌稳定HCT116细胞系24h,MTT法检测细胞增殖情况,Western blot检测自噬标志物LC3-Ⅱ的变化,并检测癌基因YAP表达变化.结果 MTT法检测结果 显示,2种药物联合应用后,肿瘤细胞的增殖抑制率明显高于单用药组(P＜0.01).姜黄素组、5-FU组和药物联用组HCT116细胞的YAP蛋白表达水平下降,LC3-Ⅱ蛋白的表达水平增高.结论 姜黄素和5-FU联用会下调YAP的表达,并诱导结肠癌细胞自噬的表达,YAP或许能成为结肠癌治疗的一个新的靶点.

Cisplatin is ranked as one of the most powerful and commonly prescribed anti-tumor chemotherapeutic agents which improve survival in many solid tumors including non-small cell lung cancer. However, the treatment of advanced lung cancer is restricted due to chemotherapy resistance. Here, we developed and investigated survivin promoter regulating conditionally replicating adenovirus (CRAd) for its anti-tumor potential alone or in combination with cisplatin in two lung cancer cells, H23, H2126, and their resistant cells, H23/CPR, H2126/CPR. To measure the expression of genes which regulate resistance, adenoviral transduction, metastasis, and apoptosis in cancer cells, RT-PCR and Western blotting were performed. The anti-tumor efficacy of the treatments was evaluated through flow cytometry, MTT and transwell assays. This study demonstrated that co-treatment with cisplatin and CRAd exerts synergistic anti-tumor effects on chemotherapy sensitive lung cancer cells and monotherapy of CRAd could be a practical approach to deal with chemotherapy resistance. Combined treatment induced stronger apoptosis by suppressing the anti-apoptotic molecule Bcl-2, and reversed epithelial to mesenchymal transition. In conclusion, cisplatin synergistically increased the tumor-killing of CRAd by (1) increasing CRAd transduction via enhanced CAR expression and (2) increasing p53 dependent or independent apoptosis of lung cancer cell lines. Also, CRAd alone proved to be a very efficient anti-tumor agent in cancer cells resistant to cisplatin owing to upregulated CAR levels. In an exciting outcome, we have revealed novel therapeutic opportunities to exploit intrinsic and acquired resistance to enhance the therapeutic index of anti-tumor treatment in lung cancer.