The expressional level of tankyrase-1 gene and its regulation in colorectal cancer in a Saudi population

Tankyrase1 plays an essential role in cancer progression by regulating telomere length. The study aimedto determine expression of TNKS1 and its regulation in colorectal cancer (CRC) in 20 samples from Saudi patients.mRNA expression of TNKS1 in CRC and paired normal tissues was measured by qRT-PCR. Epigenetic modificationof TNKS1 promoter was determined by methylation-specific PCR while somatic mutation was analyzed by Sangersequencing in exon 10 of the gene. All cancerous and normal tissues expressed TNKS1, but level of expression in CRCtissues was significantly associated with tumor stage though no other parameters; age, gender, and tumor location,showed any correlation. Expression of TNKS1 was markedly higher in earlier (I, II) than later (III, IV) stages of CRCdevelopment. Both cancerous and healthy tissues had unmethylated promoter. Sanger sequencing of exon 10 maskedany somatic mutation in the samples. Our findings suggest that up-regulation of TNKS1 was inversely correlated withcancer progression in CRC, indicating that TNKS1 participates in the initiation of CRC by stabilizing telomere length inthe first phase of cancer progression. Mechanisms other than TNKS1 might play a role in malignant tumor progressionand telomere maintenance in the late stages of CRC.     

3-<i>epi</i>-bufotalin suppresses the proliferation in colorectal cancer cells through the inhibition of the JAK1/STAT3 signaling pathway

Traditional Chinese medicine (TCM) has been increasingly employed in the last decades in China for both preventing and treating a variety of cancers. 3-epi-bufotalin is an active ingredient of TCM “Chanpi” with anti-tumor potential. However, the effect and mechanism of 3-epi-bufotalin on colorectal cancers were not well disclosed. The present study demonstrated that 3-epi-bufotalin could reduce viability, trigger apoptosis, and block the cell cycle at the G₂/M stage in colorectal cancer cell lines HT29, RKO, and COLO205 in vitro. Moreover, 3-epi-bufotalin inhibited the JAK1/STAT3 signaling pathway. These results indicated the anti-proliferation ability of 3-epi-bufotalin in colorectal cancer cells.     

Metformin and colorectal cancer

Colorectal cancer (CRC) is one of the main causes of cancer-related mortality in the developed world despite recent developments in detection and treatment. Several epidemiological studies indicate that metformin, a widely prescribed antidiabetic drug, exerts a protective effect on different cancers including CRC. Furthermore, a recent double-blind placebo-controlled, randomized trial showed that metformin significantly decreased colorectal adenoma recurrence. Studies exploring the mechanism of action of metformin in cells derived from different types of cancers reported many effects including respiratory chain complex 1 inhibition, Akt phosphorylation inhibition, ATP depletion, PKA activation and Wnt signaling inhibition. However, many of these results were obtained employing metformin at concentrations several fold higher than those achieved in target tissues in diabetic patients receiving therapeutic recommended doses of metformin. In contrast, recent studies obtained with metformin at concentrations compatible with those detected in human intestines revealed that metformin elicit responses that target β-catenin, PI3K/Akt, E-cadherin, p120-catenin and focal adhesion kinase which are key molecules and signaling pathways associated to colorectal cancer development. This brief review revisit several know aspects as well as novel ones on the effects of metformin on cancer cells.     

Therapeutic application of mesenchymal stem cells-derived extracellular vesicles in colorectal cancer

Colorectal cancer (CRC) is the third most common cancer and the leading cause of cancer death globally. Resistance to therapy is a challenge for CRC treatment. Mesenchymal stem cells (MSCs) have become one of the furthermost effective approaches for tumor treatment due to their specific feature; however, their therapeutic function is controversial. Recently, extracellular vesicles (EVs) derived from MSCs (MSCs-EVs) have attracted extensive research attention due to their promising role in CRC treatment. EVs are cell-derived vesicles that transfer different biomolecules between cells, contributing to intracellular communication. MSCs-EVs can suppress CRC by delivering therapeutic agents to tumor cells. Several studies indicate that MSCs-EVs can serve as a drug delivery system for the treatment of different cancers. Various methods are used to modify (engineer) MSCs-EVs for loading therapeutic agents. Modified MSCs-EVs have improved specificity, targeting ability, and immunogenicity compared to synthetic carriers. Furthermore, CRC-EVs participate in regulating different cells, such as immune cells, fibroblasts, and endothelial cells, promoting tumorigenesis. MSCs-EVs-based therapy indicates a high potential in the treatment of cancer; however, the majority of studies have been conducted in the pre-clinical, and their clinical applications need further scrutiny. In this review, we describe the biogenesis of EVs, focusing on the effect of MSCs-EVs on CRC cells and CRC-derived EVs on other cells. Furthermore, MSCs-EVs as a drug delivery system for cancers is also reviewed, and perspectives regarding the therapeutic application of MSCs-EVs are discussed.     

M1 macrophage-derived exosomes moderate the differentiation of bone marrow mesenchymal stem cells

Differentiated macrophages have been proven to participate in the development of mesenchymal stem cells in different tissues. However, the regulatory processes remain obscure. Exosomes, which are key secretions of macrophages, have attracted increasing attention. Therefore, macrophage-derived exosomes may modulate the development of Bone marrow mesenchymal stem cells (BMMSCs). Different culture conditions were used to induce M1 polarization in THP1 cells. Subsequently, exosomes derived from unpolarized (M0) and polarized (M1) macrophages were isolated, BMMSCs were cultured with normal complete medium or inductive medium supplemented with M0 or M1 derived exosomes, and the osteogenic capacity of the BMMSCs was measured and analyzed. Finally, molecular mechanism associated with Akt and RUNX2 was investigated. Alizarin red staining and WB experiments showed that M1 macrophages could promote the osteogenic differentiation of BMMSCs better than M0 macrophages. Then, exosomes derived from M0 and M1 macrophages were successfully isolated and analyzed by electron microscopy and WB experiments. We concluded that media containing M1-derived exosomes promoted the osteogenic differentiation of BMMSCs better than media containing M0-derived exosomes. In addition, M1-derived exosomes could activate Akt and increase RUNX2 levels to promote osteogenesis. Our data demonstrated that exosomes derived from M1 macrophages induced osteogenesis by activating Akt and increasing RUNX2 level.     

Calcium supplementation in colorectal cancer prevention: A systematic meta-analysis of adverse events

Despite the multiple systematic reviews and meta-analyses accumulating evidence on the preventive effect of calcium supplementation for colorectal cancer, most of the associated adverse effects are not systematically analyzed. The aim of the study is evaluating adverse events associated with calcium supplementation for colorectal cancer prevention through a systematic meta-analysis. We searched Medline, PubMed Central, EMBASE (Excerpta Medica database), Scopus, Cochrane Central Register of Controlled Trials, and Web of Science published in English from database inception up to 31 July 2019. In the current systematic meta-analysis, we included human studies (including cohort studies, clinical trials, case-control studies) on supplementation of calcium in patients with or at risk of colorectal cancer. Assessment of the quality of included studies was performed by Jadad score. Information on the patient population, number of enrolled subjects in each group, dose of calcium supplementation, duration of calcium supplementation, and reported adverse events were gathered. The data were pooled for incidence rates for any adverse event during the study period regardless of causality association. We identified 6 studies, comprising 4583 participants that met the inclusion criteria. Meta-analysis on pooled incidence rates for adverse event during study period showed no statistically significant increased risk for cancer (OR = 0.92, 95% CI: 0.70–1.21, P = 0.577; I² = 0.0%, P = 0.731), coronary revascularization (OR = 1.12, 95% CI: 0.79–1.59, P = 0.492; I² = 0.0%, P = 0.957), myocardial infarction (OR = 0.81, 95% CI: 0.34–1.91, P = 0.634; I² = 67.9%, P = 0.047), stroke (OR = 0.75, 95% CI: 0.42–1.33, P = 0.332, I² = 0.00%, P = 0.717), Transient Ischemic Attack (TIA) (OR = 1.37, 95% CI: 0.28–6.51, P = 0.692, I² = 81.9%, P = 0.002), urolithiasis (OR = 1.23, 95% CI: 0.75–2.01, P = 0.410; I² = 0.0%, P = 0.851), fracture (OR = 0.98, 95% CI: 0.70–1.37, P = 0.938; I² = 37.8%, P = 0.152) and death (OR = 1.05, 95% CI: 0.71–1.56, P = 0.786, I² = 12.2%, P = 0.317) in patients receiving calcium supplementation for colorectal cancer prevention compared to control. Based on the results of Egger test, publication bias was not observed among the studies (P = 0.262). The current result of the meta-analysis on human studies reporting adverse events associated with calcium supplementation for the prevention of colorectal cancer demonstrated no statistically significant increased risk for the development of adverse events compared to control groups.     

Raloxifene-loaded and aptamer-bonded exosomes induce autophagic and apoptotic death in HeLa cells by enhancing the lysosomotropic effect

Background: Raloxifene, a selective estrogen receptor modulator, is also known to be a lysosomotropic agent. The bioavailability of raloxifene is around 2% due to extensive hepatic transport. Exosomes are nanosized vesicles that are naturally released from cells. Method: In this study, exosomes released from HeLa cervical cancer cells were loaded with raloxifene to increase its bioavailability, and an aptamer was attached to the exosome membrane for targeting only HeLa cells. Characterization of exosomes isolated from HeLa cells was performed by transmission electron microscopy, zeta sizer, and western blotting. In addition, the cytotoxic, apoptotic, autophagic, and lysosomotropic effects of the prepared Exo-Apt-Ral formulation on HeLa cervical cancer cells were investigated. Results: According to zeta analysis, the sizes of the empty exosome and Exo-Apt-Ral formulation were measured as 66 ± 12 and 120 ± 21 nm, respectively. There was a rise in the lysosomal permeability of HeLa cells after the Exo-Apt-Ral application. In addition, both apoptotic and autophagic death mechanisms were triggered in HeLa cells after the Exo-Apt-Ral application. Conclusion: This study showed that raloxifene functionalized by loading into aptamer-bound exosomes can be a new targeted drug carrier system for cervical cancer.     

Interaction of IL-22/IL-22R1 promotes cell proliferation and suppresses apoptosis of colorectal cancer via phosphorylation of STAT3

Interleukin-22 (IL-22) is a member of IL-10 cytokine family which is expressed in activated T cellspredominantly and in activated natural killer cells at lower levels. Previous studies have demonstrated the link betweenelevated levels of IL-22 and disease severity of psoriasis, Crohn’s disease, rheumatoid arthritis and interstitial lungdiseases. However, the function of IL-22 in the development and progression of colorectal cancer (CRC) remainselusive. In this study, we first evaluated the IL-22/IL-22R1 level in CRC patients, and found that tumor tissueshave more active expression of IL-22 and IL-22R1 than normal tissues, presenting correlation with the degree ofdifferentiation of tumor tissues. Subsequently, Caspase and cell viability assays were performed on SW-480 cell linewhich expresses high level of IL-22R1 to examine if the supplementation of IL-22 has an impact on apoptosis andproliferation. In comparison with treatment of 5-FU, supplementation of IL-22 promoted cell proliferation andameliorated apoptosis. To unveil signal transduction upon activation of IL-22R, we examined the phosphorylationof STAT3 in SW-480 cell line following supplementation of IL-22. The treatment of IL-22 also increased the level ofp-Akt, an essential component in PI3K/Akt pathway. Although the link between STAT3 phosphorylation and PI3K/Akt activation remains to be explored, our study revealed the mechanism underlying the effects of IL-22R activation onapoptosis as well as tumor differentiation, indicating the prognostic value of IL-22/IL-22R.     

Elp3 modulates neural crest and colorectal cancer migration requiring functional integrity of HAT and SAM domains

Cell migration is a finely tuned biological process that often involves epithelial-mesenchymal transition (EMT).EMT is typically characterized by the upregulation of mesenchymal markers such as Snail1. This process has been shownto be of critical importance to normal developmental processes, including neural crest migration and invasion.Interestingly, similar mechanisms are utilized in disease processes, such as tumor metastasis and migration. Notably,EMT and EMT-like processes confer tumor cells with the ability to migrate, invade, and adopt stem cell-likeproperties that largely account for immunosuppression and tumor recurrence. Therefore, identifying sensitive EMTmarkers may contribute to cancer prognosis and diagnosis in many facets. Previously, we showed that Elp3 plays anessential role during neural crest migration by stabilizing Snail1. In the current study, we further elucidate themolecular mechanism underlying colorectal cancer migration. We found that mElp3 exerted an identical function toxElp3 in modulating neural crest migration, and both HAT and SAM domains are imperative during this migratoryprocess. Interestingly, overexpression of mElp3 in SW480 cells promoted cell migration and invasion, and we furthershowed that Elp3 stabilized Snail1 requiring integrity of both SAM and HAT domains. Our findings warrant furtherexploration of the specific target of Elp3 in cancer cells.     

Knockdown Wiskott-Aldrich syndrome protein family member 3 (WASF3) inhibits colorectal cancer metastasis and sensitizes to cisplatin through targeting ZNF471

Colorectal cancer (CRC) is a heterogeneous cancer, and many risk factors for colorectal cancer have beenestablished. For CRC metastasis, tumor cell migration, adhesion as well as invasion are important processes. WiskottAldrich syndrome protein family member 3 (WASF3) is necessary for metastasis of various types of cancers. However,its role in CRC progression has not been fully elucidated. This study examined the in vitro functional roles of WASF3 inthe CRC and explored the underlying molecular mechanisms. We used siRNA-WASF3 to gene silence WASF3 in coloncancer cell (HCT116) in vitro. The effects of WASF3 silencing on HCT116 cell apoptosis, proliferation, migration, aswell as invasion were assessed by flow cytometry, CCK-8, and transwell assays. ZNF471 protein expressions weredetected by immunofluorescence staining and RT-PCR. Moreover, the effects of ZNF471 were studied on a series of invitro antitumor-promoting assays using HCT116. WASF3 knockdown expression using small interfering RNA (siRNA)ameliorated CRC cell proliferation, anchorage-independent growth, invasion, and metastasis. Furthermore, we observedthat WASF3 contributed to upregulating the metastasis signaling pathway through inhibiting the expression of ZNF471.Our study suggests that targeting WASF3 signaling might be a novel therapeutic strategy for treating CRC.     

Effect of microRNA-143-3p-mediated CTNND1 on the biological function of lung cancer cells

Lung cancer poses a serious threat to human life with high incidence and miRNA is an important biomarkerin tumors. This study aimed to explore the effect of miR-143-3p on the biological function of lung cancer cells and theunderlying mechanism. Eighty-seven samples of lung cancer tissues and 81 samples of tumor-adjacent tissues frompatients undergoing radical lung cancer surgery in our hospital were collected. The lung cancer cells and lung fibroblastcells (HFL-1) were purchased, and then miR-143-3p-mimics, miR-NC, si-CTNND1, and NC were transfected intoA549 and PC-9 cells to establish cell models. MiR-143-3p and CTNND1 expression levels were measured by the qRTPCR,Bax, Bcl-2, and CTNND1 expression levels by the Western Blot (WB), and cell proliferation, invasion, andapoptosis by the MTT assay, Transwell assay, and flow cytometry. Dual luciferase report assay was used to determinethe relationship between miR-143-3p and CTNND1. In this study, miR-143-3p was lowly expressed in lung cancerand CTNND1 was highly expressed in lung cancer. The overexpression of miR-143-3p inhibited cell proliferationand invasion, promoted cell apoptosis, significantly increased Bax protein expression, and decreased Bcl-2 proteinexpression. The inhibition of CTNND1 led to opposite biological characteristic in cells. The dual luciferase reporterassay demonstrated that miR-143-3p was a target region of CTNND1. Such results suggest that miR-143-3p can inhibitthe proliferation and invasion of lung cancer cells by regulating the expression of CTNND1 and promote the apoptosisof lung cancer cells, sott is expected to be a potential target for lung cancer.     

Her-2与ALDH1在乳腺癌中表达与相关性研究

目的:探讨乳腺癌组织中Her-2及ALDH1的表达相关性及其临床意义。方法:选取105例原发性乳腺癌标本。采用免疫组化法检测Her-2、ALDH1蛋白的表达,并结合临床病理特征进行相关性分析和无病生存期分析。结果:(1)Her-2与ALDH1在乳腺癌中的表达率分别为29.52%、66.67%。(2)乳腺癌原发灶中Her-2蛋白的表达与肿瘤大小、ER呈正相关(P≤0.05),与年龄、民族、组织学分期及病理类型、PR均无相关性(P>0.05)。ALDH1蛋白表达与年龄、民族、组织学分期及病理类型、肿瘤大小、淋巴结转移、ER、PR均未发现有相关性(P>0.05)。(3)Her-2与ALDH1在乳腺癌表达中呈正相关(r=0.192,P≤0.05)。(4)Her-2高表达患者2年无病生存率显著低于Her-2阴性患者(P<0.05),ALDH1阳性患者与阴性患者2年无病生存期无明显差异(P>0.05)。(5)COX回归模型分析结果显示Her-2是影响患者预后的独立因素(P=0.008)。Her-2的相对危险度=6.284>1。结论:ALDH1与Her-2在乳腺癌组织中的表达呈正相关。Her-2表达与乳腺癌术后患者预后呈负相关,而ALDH1表达与乳腺癌术后患者的预后无明显相关性。     

Analysis of tumor-draining vein secretome: A direct access to tumor-derived extracellular vesicles in surgical lung cancer patients

Tumor-secreted extracellular vesicles (EVs) participate in the metastasis process through different mechanisms, including the preparation of the pre-metastatic niche to grant circulating tumor cells (CTCs) implantation and growth. The study of the metastasis process through the analysis of CTCs and tumor-derived EVs is difficult because of the dilution grade of these elements in peripheral blood. In early-stage lung cancer patients, the tumor-secreted products are even more diluted. An attractive strategy in surgical lung cancer patients is to purify them from a pulmonary tumor-draining vein where they are enriched. The information obtained from the analysis of EVs and CTCs purified from this source could give more accurate information about tumor biology and could be an important source of biomarkers to identify patients at high risk of relapse after curative surgery.     

<i>XRCC1</i> Arg399Gln and Arg194Trp polymorphisms regulate XRCC1 expression and chemoresistance of non-small cell lung cancer cells

X-ray repair cross-complementing protein 1 (XRCC1) could repair cisplatin-induced DNA damage. XRCC1Arg399Gln and Arg194Trp variants alter XRCC1 expression and function, leading to changes in cancer sensitivity tocisplatin treatment. This study aimed to investigate the effects of XRCC1 Arg399Gln and Arg194Trp polymorphismson cell viability, apoptosis and XRCC1 expression in cisplatin-sensitive A549 and cisplatin-resistant A549/DDP nonsmall cell lung cancer (NSCLC) cells. Plasmids carrying XRCC1 Arg399Gln and Arg194Trp were constructed andtransfected into A549 and A549/DDP cells. RT–PCR, Western blot, MTT assay, and flow cytometry analysis wereperformed to assess cell viability, apoptosis, and XRCC1 expression. Compared to control cells, the viability of A549and A549/DDP cells transfected with XRCC1 Arg399Gln and Arg194Trp was higher and the apoptosis rate was lower,and XRCC1 mRNA and protein expression levels were significantly higher. In conclusion, our results suggest thatXRCC1 Arg399Gln and Arg194Trp polymorphisms change XRCC1 expression in NSCLC cells and alter the sensitivityof NSCLC to cisplatin-based chemotherapy     

Silencing of long non-coding RNA CCHE1 inhibits the ovarian cancer SKOV3 cell invasion and migration and inactivates the p38-MAPK signaling pathway

Ovarian cancer (OC) is a major cause of cancer-related deaths among gynaecological malignancies. Emerging studiessuggest that the long non-coding RNA (lncRNA) may be the potential biomarker for the diagnosis and prognosis of the cancer.The current study was carried out to investigate the role of lncRNA CCHE1 silencing in OC cell invasion and migration.Expression of lncRNA CCHE1 in normal ovarian cell Hose and OC cell lines HO 8910, A2780 and SKOV3 was detected.LncRNA were transfected with siRNA, and then the proliferation of cells was detected by using MTT assay. Cell invasionand migration was measured by using Transwell assay and scratch test, respectively. The protein levels of E-cadherin,N-cadherin, ERK, p38-MAPK and the phosphorylation of ERK and p38-MAPK in cells after siRNA transfection weredetected by using Western blot analysis. Consequently, lncRNA CCHE1 expression was highly expressed in OC cell lines,especially in SKOV3 cells. siRNA1, siRNA2 and siRNA3 all decreased. lncRNA CCHE1 expression in SKOV3 cells andsiRNA2 showed the best silencing efficacy. Silencing of lncRNA CCHE1 decreased proliferation, invasion and migration,and reduced the protein levels of N-cadherin, ERK, p38-MAPK and the phosphorylation of ERK and p38-MAPK, whilereducing the protein level of E-cadherin in SKOV3 cells. Collectively, our study proved that the silencing of lncRNACCHE1 could inhibit SKOV3 cell invasion and migration via inactivating the p38-MAPK signaling pathway.