Colon cancer-associated B2 Escherichia coli colonize gut mucosa and promote cell proliferation

AIM: To provide further insight into the characterization of mucosa-associated Escherichia coli (E. coli) isolated from the colonic mucosa of cancer patients.

Tetramethylpyrazine inhibits proliferation of colon cancer cells in vitro

BACKGROUND Colon cancer is one of the most common malignancies worldwide,and chemotherapy is a widely used strategy in colon cancer clinical therapy.However,chemotherapy resistance is a major cause of disease recurrence and progression in colon cancer,and thus novel drugs for treatment are urgently needed.Tetramethylpyrazine(TMP),a component of the traditional Chinese medicine Chuanxiong Hort,has been proven to exhibit a beneficial effect in tumors.AIM To investigate the potential anticancer activity of TMP in colon cancer and its underlying mechanisms.METHODS Colon cancer cells were incubated with different concentrations of TMP.Cell viability was evaluated by crystal violet staining assay and cell counting kit-8 assay,and cell apoptosis and cell cycle were assessed by flow cytometry.RESULTS TMP significantly inhibited the proliferation of colon cancer cells in a dose-and time-dependent manner.In addition,flow cytometry revealed that TMP induced cell cycle arrest at the G0/G1 phase.TMP treatment caused early stage apoptosis in SW480 cells,whereas it caused late stage apoptosis in HCT116 cells.CONCLUSION Our studies demonstrated that TMP inhibits the proliferation of colon cancer cells in a dose-and time-dependent manner by inducing apoptosis and arresting the cell cycle at the G0/G1 phase.Our findings suggest that TMP might serve as a potential novel therapeutic drug in the treatment of human colon cancer.

EFFECT OF TNF-a AND IFN-g ON THE EXPRESSION OF INDUCIBLE NITRIC OXIDE SYNTHASE GENE AND PROLIFERATION INHIBITION OF HUMAN COLON CANCER CELL LINE

Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon cancer cell line CCL229. Methods: Using the molecular and biochemical techniques and electron microscopy to analyze the expression of iNOS, production of NO and growth characteristics of human colon cancer cells. Results: cytokine treatment can induce expression of the iNOS gene and production of nitric oxide was significantly higher after treatment of CCL229 cells with TNF-αor IFN-γ. Treatment with either cytokine or a combination of both significantly increased levels of Malondialdehyde (MDA) over control. Furthermore, cytokine treatment increased the proliferation inhibition rate as assessed in vitro and decreased the cell proliferation index on flow cytometry. Electron microscopy showed that cells treated with cytokines had fewer pseudopodia or cell processes than control cells and that cytokine treated cells had dilatation of the mitochondria and endoplasmic reticulum and dilated vesicular or tubular cisternae. Conclusion: Our findings indicate that TNF-α and IFN-γ induce the expression of iNOS gene in CCL229 cells, which increases the production of nitric oxide, inhibits proliferation, causes lipid peroxidation, and results in ultrastructural changes.

Effects of MIF on proliferation,migration,and STAT1 pathway of colon cancer cells

Objective This study aimed to investigate how macrophage migration inhibitory factor(MIF)regulates the interaction of signal transducer and activator of transcription 1(STAT1)with CD74,and affects colon cancer proliferation and invasion.Methods After transfecting MIF small interfering RNA into the SW480 cell line,the expression of STAT1 and CD74 mRNA was detected by qRT-PCR and western blotting.Transwell and MTT assays were performed to detect the colon cancer cell invasion and proliferation ability.Co-immunoprecipitation was used to detect the interaction between CD74 and STAT1 proteins in the treated and control groups.Results The cellular biological assays(MTT and Transwell)showed that the proliferation and invasion ability of colon cancer cells decreased after MIF knockdown;the results showed significant statistical difference(P<0.05).The results of the co-immunoprecipitation assay suggested that MIF knockdown in colon cancer cells could inhibit the binding of CD74 and STAT1 proteins;statistical difference was observed between the two groups(P<0.05).Conclusion MIF can increase the proliferation and invasion of colon cancer cells by promoting the combination of CD74 and STAT1.

Correlation of dynamic contrast-enhanced magnetic resonance imaging parameters before colon cancer surgery with the angiogenesis and cell proliferation in tumor lesions

Objective: To discuss the correlation of dynamic contrast-enhanced magnetic resonance imaging parameters before colon cancer surgery with the angiogenesis and cell proliferation in tumor lesions. Methods: A total of 186 patients with colon cancer who were treated in the hospital between January 2015 and January 2017 were collected as colon cancer group;100 patients with multiple polyposis of colon who received colonoscopy in the hospital during the same period were selected as colon polyp group. The differences in DCE-MRI parameters as well as the expression of angiogenesis and cell proliferation genes were compared between the two groups, and Pearson test was used to evaluate the correlation of preoperative DCE-MRI parameters with angiogenesis and cell proliferation gene expression in patients with colon cancer. Results: Preoperative DCE-MRI parameters Ktrans and Kep levels in colon cancer group were significantly higher those in colon polyp group;Cyr61, HIF-α, VEGF, MMP-9, CXCR7, EZH2, SphK1 and IFT57 mRNA expression in lesion tissue of colon cancer group were significantly higher than those of colon polyp group while Kiss-1 mRNA was lower than that of colon polyp group. Pearson test showed that the DCE-MRI parameters Ktrans and Kep levels before colon cancer surgery were directly correlated with the expression of angiogenesis and cell proliferation genes in lesion tissues. Conclusion: Preoperative DCE-MRI parameters can accurately reflect the severity of colon cancer.

Stem cell characteristics of early colon cancer tissue and their relationship with cell proliferation and invasion

Objective:To study the stem cell characteristics of early colon cancer tissue and their relationship with cell proliferation and invasion.Methods: Colon cancer tissues and adjacent tissues surgically removed in Dongguan People's Hospital between January 2010 and October 2017 were selected as the clinical samples of this study, the protein was extracted to determine the protein expression of tumor stem cell genes USP22, Nanog, Lgr5 and CD44, and RNA was extracted to determine the mRNA expression of cell proliferation genes and cell invasion genes.Results:USP22, Nanog, Lgr5 and CD44 protein expression as well as Rab5A, TBX2, MDM2, TGF-β1, Smad2/3, Vimentin, Rac1 and VEGF mRNA expression in colon cancer tissues were significantly higher than those in adjacent tissues while Bad, Bax and Fas mRNA expression were significantly lower than those in adjacent tissues;USP22, Nanog, Lgr5 and CD44 protein expression in colon cancer tissues were positively correlated with Rab5A, TBX2, MDM2, TGF-β1, Smad2/3, Vimentin, Rac1 and VEGF mRNA expression, and negatively correlated with Bad, Bax and Fas mRNA expression.Conclusion: The activation of stem cell characteristics in early colon cancer can promote the proliferation and invasion of cancer cells.

Effect of nutritional support + intravenous chemotherapy on anti-tumor immunity and cancer cell proliferation in patients with colon cancer complicated by incomplete intestinal obstruction

Objective:To study the effect of nutritional support + intravenous chemotherapy on anti-tumor immunity and cancer cell proliferation in patients with colon cancer complicated by incomplete intestinal obstruction.Methods: Patients with colon cancer complicated by incomplete intestinal obstruction who were treated in Midi Branch, Pangang Group General Hospital between March 2015 and October 2017 were selected and randomly divided into the nutrition group who accepted nutritional support + FOLFOX4 intravenous chemotherapy and the control group who accepted FOLFOX4 intravenous chemotherapy alone, and they underwent surgery after two cycles of chemotherapy. The contents of immune cells in peripheral blood and the contents of immune cytokines in serum were determined before chemotherapy and two cycles after chemotherapy;the expression levels of proliferation genes in colon cancer lesions were determined after surgical resection.Results:Compared with those of same group before chemotherapy, peripheral blood Treg, Th9, Th17 and Th22 contents as well as serum IL-4, IL-9, IL-10, TGF-β1, IL-17 and IL-22 contents of nutrition group were decreased significantly after chemotherapywhereas peripheral blood Treg, Th9, Th17 and Th22 contents as well as serum IL-4, IL-9, IL-10, TGF-β1, IL-17 and IL-22 contents of control group did not change significantly after chemotherapy, and compared with those after chemotherapy between groups, peripheral blood Treg, Th9, Th17 and Th22 contents as well as serum IL-4, IL-9, IL-10, TGF-β1, IL-17 and IL-22 contents of nutrition group were significantly lower than those of control group, and CyclinD1, Bcl-2, USP22, VEGF and N-cadherin mRNA expression were not different from those of control group.Conclusion:Nutritional support + intravenous chemotherapy can improve the anti-tumor immune response without affecting the proliferation of cancer cells in the lesion of patients with colon cancer complicated by incomplete intestinal obstruction.

Effect of soy saponin on the growth of human colon cancer cells

AIM:To investigate the effect of extracted soybean saponins on the growth of human colon cancer cells.METHODS:WiDr human colon cancer cells were treated with 150,300,600 or 1200 ppm of soy saponin to determine the effect on cell growth,cell morphology,alkaline phosphatase(AP) and protein kinase C(PKC) activities,and P53 protein,c-Fos and c-Jun gene expression.RESULTS:Soy saponin decreased the number of viable cells in a dose-dependent manner and suppressed 12-Otetradecanol-phorbol-13-acetate-stimulated PKC activity(P < 0.05).Cells treated with saponins developed cytoplasmic vesicles and the cell membrane became rougher and more irregular in a dose-dependent manner,and eventually disassembled.At 600 and 1200 ppm,the activity of AP was increased(P < 0.05).However,the apoptosis markers such as c-Jun and c-Fos were not significantly affected by saponin.CONCLUSION:Soy saponin may be effective in preventing colon cancer by affecting cell morphology,cell proliferation enzymes,and cell growth.

Down-regulation of p110β Expression Increases Chemosensitivity of Colon Cancer Cell Lines to Oxaliplatin

This study examined the synergetic effect of class ⅠA Phosphoinositide 3-kinases catalytic subunit p110β knockdown in conjunction with oxaliplatin treatment on colon cancer cells. Down-regulation of p110β by siRNA interference and oxaliplatin treatment were applied in colon cancer cell lines HT29, SW620 and HCT116. MTT assay was used to measure the inhibitory effect of p110β knockdown on the proliferation of colon cancer cell lines. SubG1 assay and Annexin-Ⅴ FITC/PI double-labeling cytometry were applied to detect cell apoptosis. And cell cycle was evaluated by using PI staining and flow cytometry. The expression of caspase 3, cleaved PARP, p-Akt, T-Akt and p110β was determined by western blotting. The results suggested that down-regulation of p110β expression by siRNA obviously reduced cell number via accumulation in G0-G1 phase of the cell cycle in the absence of notablely increased apoptosis in colon cancer cell lines HT29 and SW620 (S phase arrest in HCT116). Moreover, inhibition of p110β expression increased oxaliplatin-induced cell apoptosis and cell cycle arrest in HT29, HCT116 and SW620 cell lines. In addition, increases of cleaved caspase-3 and cleaved PARP induced by oxaliplatin treatment were determined by immunoblotting in p110β knockdown group compared with normal control group and wildtype group. It is concluded that down-regulated expression of p110β could inhibit colon cancer cells proliferation and result in increased chemosensitivity of colorectal cancer cells to oxaliplatin through augmentation of oxaliplatin-induced cell apoptosis and cell cycle arrest.

Growth inhibition of colon cancer cells by compounds affecting AMPK activity

AIM:To determine if other molecules reported to modulate AMP-dependent protein kinase(AMPK)activ-ity would have effects resembling those of metformin and phenformin on colon cancer cell proliferation and metabolism.METHODS:Studies were performed with four hu-man colon cancer cell lines,Caco-2,HCT116,HT29 and SW1116.The compounds that were studied included A-769662,5-aminoimidazole-4-carboxamide-1-ribofu-ranoside,butyrate,(-)-epigallocatechin gallate(EGCG),KU-55933,quercetin,resveratrol and salicylates.The parameters that were measured were cell proliferation and viability,glucose uptake,lactate production and acidification of the incubation medium.RESULTS:Investigations with several molecules that have been reported to be associated with AMPK activa-tion(A-769662,5-aminoimidazole-4-carboxamide-1-b-D-ribofuranoside,EGCG,KU-55933,quercetin,resve-ratrol and salicylates)or AMPK inhibition(compound C)failed to reveal increased medium acidification and increased glucose uptake in colon cancer cells as previ-ously established with metformin and phenformin.The only exception was 5-aminosalicylic acid with which there were apparently lower glucose levels in the me-dium after incubation for 72 h.Further study in the absence of cells revealed that the effect was an artifact due to inhibition of the enzyme-linked glucose assay.The compounds were studied at concentrations that inhibited cell proliferation.CONCLUSION:It was concluded that treatment with several agents that can affect AMPK activity resulted in the inhibition of the proliferation of colon cancer cells under conditions in which glucose metabolism is not enhanced,in contrast to the effect of biguanides.

Inhibition of self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells by scutellarin via Hedgehog signaling pathway

OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29CSC)in vitro and in vivo,and to explore its mechanism.METHODS The effect of scutellarin on the growth of HT-29CSC was determined by 3D Culture assay.The effect of scutellarin on growth and transformation of HT-29CSC was probed by soft agar colony formation assay.The effect of scutellarin on the differentiation of HT-29CSC was determined by serum induction differentiation assay in vitro.The effects of scutellarin on the expressions of marker gene Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog gene were measured by quantitative real-time RT-PCR.Investigate the effect of scutellarin on the expression of c-Myc,Gli1,and Lgr5 protein by Western blotting.A subcutaneous xenograft model of colon cancer in nude mice was established and administered by intraperitoneal injection.The change of body weight and tumor size of nude mice were observed every two days.Investi⁃gate the effects of scutellarin on the growth of xenograft tumors in nude mice.The expression of CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,Nanog gene in tumors were measured by quantitative real-time RT-PCR.The expression of c-Myc,Gli1,Lgr5,CD133,Ki67 protein were measured by Western blotting.RESULTS Scutellarin can inhibit the growth of HT-29CSC in 3D culture.Compared with the solvent control group,scutellarin can significantly inhibit the growth and transformation and differentiation of HT-29CSC in vitro(P<0.01).The expression levels of marker genes Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog in HT-29CSC were down-regulated by scutellarin.Scutellarin can reduce the expression of c-Myc,Gli1,and Lgr5 protein in HT-29CSC.Scutellarin can inhibit the growth of colon cancer xenografts,lower CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,and Nanog mRNA level of xenograft tumors,reduce the expression of c-Myc,Gli1,Lgr5,CD133,and Ki67 protein of xenograft tumors in nude mice.CONCLUSION Scutellarin,which is the main component of scutellaria barbata,can inhibit the differentiation of HT-29CSC and the mechanism is to inhibit the activity of Hedgehog signaling pathway.

Isolation and identification of adipose-derived stromal/stem cells from breast cancer patients after exposure neoadjuvant chemotherapy

BACKGROUND With recent research advances,adipose-derived stromal/stem cells(ASCs)have been demonstrated to facilitate the survival of fat grafts and thus are increasingly used for reconstructive procedures following surgery for breast cancer.Unfortunately,in patients,following radiation and chemotherapy for breast cancer suggest that these cancer treatment therapies may limit stem cell cellular functions important for soft tissue wound healing.For clinical translation to patients that have undergone cancer treatment,it is necessary to understand the effects of these therapies on the ASC's ability to improve fat graft survival in clinical practice.AIM To investigate whether the impact on ASCs function capacity and recovery in cancer patients may be due to the chemotherapy.METHODS ASCs were isolated from the cancerous side and noncancerous side of the breast from the same patients with receiving neoadjuvant chemotherapy(NAC)or notreceiving NAC.ASCs were in vitro treated with 5-fluorouracil(5-FU),doxorubicin(DXR),and cyclophosphamide(Cytoxan)at various concentrations.The stem cells yield,cell viability,and proliferation rates were measured by growth curves and MTT assays.Differentiation capacity for adipogenesis was determined by qPCR analysis of the specific gene markers and histological staining.RESULTS No significant differences were observed between the yield of ASCs in patients receiving NAC treatment and not-receiving NAC.ASCs yield from the cancerous side of the breast showed lower than the noncancerous side of the breast in both patients receiving NAC and not-receiving NAC.The proliferation rates of ASCs from patients didn’t differ much before and after NAC upon in vitro culture,and these cells appeared to retain the capacity to acquire adipocyte traits simile to the ASCs from patients not-receiving NAC.After cessation and washout of the drugs for another a week of culturing,ASCs showed a slow recovery of cell growth capacity in 5-FU-treated groups but was not observed in ASCs treated with DXR groups.CONCLUSION Neoadjuvant therapies do not affect the functioning capacity of ASCs.ASCs may hold great potential to serve as a cell source for fat grafting and reconstruction in patients undergoing chemotherapy.

Upregulation of 25-hydroxyvitamin D_3-1α-hydroxylase by butyrate in Caco-2 cells

AIM： To investigate the possible involvement of 25-hydroxyvitamin D3-1cx-hydroxylase [1α-25（OH）2D3] in butyrate-induced differentiation in human intestinal cell line Caco-2 cells. METHODS： Caco-2 cells were incubated either with 3 mmol/L butyrate and 1 umol/L 25（OH）2D3 or with 1 umol/L 1α-25（OH）2D3 for various time intervals ranging from 0 to 72 h. Additionally, cells were co-incubated with butyrate and either 25（OH）2D3 or 1α-25（OH）2D3. 1α-25（OH）2D3 mRNA was determined semi-quantitatively using the fluorescent dye PicoGreen. Immunoblotting was used for the detection of 1α-25（OH）2D3 protein. Finally, enzymatic activity was measured by ELISA. RESULTS： Both butyrate and 1α-25（OH）2D3 stimulated differentiation of Caco-2 cells after a 48 h incubation period, while 25（OH）2D3 had no impact on cell differentiation. Synergistic effects on differentiation were observed when cells were co-incubated with butyrate and vitamin D metabolite. Butyrate transiently upregulated 1α-25（OH）2D3 mRNA followed by a timely delayed protein upregulation. Coincidently, enzymatic activity was enhanced significantly. The induction of the enzyme allowed for comparable differentiating effects of both vitamin D metabolites. CONCLUSION： Our experimental data provide a further mechanism for the involvement of the vitamin D signaling pathway in colonic epithelial cell differentiation by butyrate. The enhancement of 1α-25（OH）2D3 followed by antiproliferative effects of the vitamin D prohormone in the Caco-2 cell line suggest that 25（OH）2D3 in combination with butyrate may offer a new therapeutic approach forthe treatment of colon cancer.

Minibrain-related kinase/dual-specificity tyrosine-regulated kinase 1B implication in stem/cancer stem cells biology

Dual-specificity tyrosine phosphorylation-regulated kinase 1B(DYRK1B),also known as minibrain-related kinase(MIRK)is one of the best functionally studied members of the DYRK kinase family.DYRKs comprise a family of protein kinases that are emerging modulators of signal transduction pathways,cell proliferation and differentiation,survival,and cell motility.DYRKs were found to participate in several signaling pathways critical for development and cell homeostasis.In this review,we focus on the DYRK1B protein kinase from a functional point of view concerning the signaling pathways through which DYRK1B exerts its cell type-dependent function in a positive or negative manner,in development and human diseases.In particular,we focus on the physiological role of DYRK1B in behavior of stem cells in myogenesis,adipogenesis,spermatogenesis and neurogenesis,as well as in its pathological implication in cancer and metabolic syndrome.Thus,understanding of the molecular mechanisms that regulate signaling pathways is of high importance.Recent studies have identified a close regulatory connection between DYRK1B and the hedgehog(HH)signaling pathway.Here,we aim to bring together what is known about the functional integration and cross-talk between DYRK1B and several signaling pathways,such as HH,RAS and PI3K/mTOR/AKT,as well as how this might affect cellular and molecular processes in development,physiology,and pathology.Thus,this review summarizes the major known functions of DYRK1B kinase,as well as the mechanisms by which DYRK1B exerts its functions in development and human diseases focusing on the homeostasis of stem and cancer stem cells.

Correlation of serum GDF-15 and MIF contents with the malignant behaviors of cancer cells in patients with NSCLC

Objective:To study the correlation of serum GDF-15 and MIF contents with the malignant behaviors of cancer cells in patients with NSCLC.Methods: Patients with non-small cell lung cancer who underwent surgical resection in Kashgar Prefecture First People's Hospital in the Xinjiang Uygur Autonomous Region between January 2015 and November 2017 were selected as lung cancer group for the research, and healthy volunteers who received physical examination in Kashgar Prefecture First People's Hospital in the Xinjiang Uygur Autonomous Region during the same period were selected as the control group. Serum was collected from the lung cancer group before surgery and from the control group during physical examination respectively to determine the contents of GDF-15 and MIF;lung cancer tissue and adjacent tissue were collected from lung cancer group after surgery to determine the expression of tumor suppressor genes, proliferation genes and invasion genes.Results:Serum GDF-15 and MIF contents of lung cancer group were significantly higher than those of control group;TCF21, Bax, GRPC5A and PTEN mRNA expression in lung cancer tissue were significantly lower than those in the adjacent tissue whereas Bcl-2, AQP4, c-myc, CyclinD1, SIRT1, CatL, MMP9, N-cadherin and Vimentin mRNA expression were significantly higher than those in adjacent tissue;serum GDF-15 and MIF contents of patients with lung cancer were negatively correlated with TCF21, Bax, GRPC5A and PTEN mRNA expression, and positively correlated with Bcl-2, AQP4, c-myc, CyclinD1, SIRT1, CatL, MMP9, N-cadherin and Vimentin mRNA expression in lung cancer tissue.Conclusion:The abnormal increase of GDF-15 and MIF in patients with NSCLC is closely related to the abnormal proliferation and invasion of cancer cells.

沉默E26转录因子变异体4抑制核因子-κB信号通路对结肠癌细胞增殖和迁移的影响

目的探讨E26转录因子变异体4(ETV4)通过核因子-κB(NF-κB)信号通路对结肠癌细胞增殖和迁移的影响机制。方法通过用户友好的交互式癌症转录组数据分析资源(UALCAN)数据库分析ETV4在结肠正常组织和癌组织中的表达;采用逆转录定量聚合酶链式反应(qRT-PCR)和Western blot检测ETV4在正常肠上皮细胞和结肠癌细胞系中的表达;沉默SW480细胞的ETV4后,采用qRT-PCR和Western blot检测ETV4的表达以评估转染效率;采用菌落形成实验和Transwell实验检测沉默ETV4后对结肠癌细胞增殖和迁移的影响;采用Western blot检测沉默ETV4后对NF-κB通路中的蛋白65(p65)和磷酸化蛋白65(p-p65)的蛋白表达的影响。结果UALCAN数据库分析结果显示ETV4在结肠癌组织中高表达。qRT-PCR和Western blot检测显示ETV4在结肠癌细胞系SW480、Lovo、Caco-2和SW620中的表达高于正常肠上皮细胞HIEC-6,其中SW480细胞中ETV4的表达最高,差异有统计学意义(P<0.001)。菌落形成实验和Transwell实验结果显示,沉默ETV4后显著抑制了结肠癌细胞SW480的增殖和迁移能力(P<0.001)。Western blot检测结果显示,沉默ETV4显著抑制细胞中p-p65蛋白的表达(P<0.001)。结论沉默ETV4可能抑制NF-κB信号通路的激活,进而抑制结肠癌细胞的增殖和迁移。

线粒体核糖体蛋白S35对结肠癌细胞增殖、侵袭和迁移的调控作用及机制研究

目的探讨线粒体核糖体蛋白S35(MRPS35)对结肠癌细胞增殖、侵袭和迁移的调控作用及机制。方法收集120例结肠癌根治术患者的结肠癌组织及癌旁正常组织标本,培养人结肠癌细胞系(HCT116、SW480、SW620)和人正常结肠上皮细胞系(NCM460)。通过生物信息学分析、实时荧光定量聚合酶链反应、蛋白质印迹法(Western blot)、免疫组织化学(IHC)分析、细胞功能实验(平板克隆形成实验、划痕实验、Transwell细胞迁移实验、CCK-8细胞活力实验)等方法评估MRPS35在结肠癌中的表达及调控机制。结果生物信息学分析结果显示,MRPS35基因在结直肠癌组织中的表达水平高于癌旁正常组织,差异有统计学意义(P<0.05)。人结肠癌细胞系(HCT116、SW480、SW620)中MRPS35 mRNA和MRPS35蛋白相对表达量均高于NCM460细胞,差异有统计学意义(P<0.05);结肠癌组织中的MRPS35蛋白相对表达量高于癌旁正常组织,差异有统计学意义(P<0.05)。MRPS35表达水平与肿瘤直径、肿瘤分化程度、T分期显著相关(P=0.002、0.021、0.036)。MRPS35高表达患者的总体生存率高于MRPS35低表达患者,差异有统计学意义(Log-rank P=0.015)。敲低MRPS35后,结肠癌细胞克隆、增殖、侵袭、迁移能力均显著增强。敲低MRPS35后,Wnt1、β-Catenin及其下游靶标蛋白的表达显著增加。结论MRPS35在结肠癌组织和结肠癌细胞中均显著高表达,其可能通过调控Wnt/β-Catenin信号通路抑制结肠癌的发生与发展,有望成为结肠癌的新型生物标志物和潜在治疗靶点。

Integrin α6β4 in colorectal cancer

The ability of cells to interact with extracellular matrix macromolecules is at the forefront of the regulation of cell phenotype and organization.Indeed most if not all cells bear specific cell surface receptors for these molecules,namely the integrins,which are specific for the ligation of various macromolecules such as the laminins,fibronectins and tenascins.It is now well established that integrins can regulate a variety of biological activities,most notably cell cycle and tissue-specific gene expression.In the intestine,several observations suggest functional roles for cell-matrix interactions in the regulation of epithelial cell functions.This article focuses on integrin α6β4 as a paradigm to illustrate the importance as well as the complexity of integrins in the mediation of cell-matrix interactions.Indeed,α6β4 has been well-characterized for its involvement as a link between the cytoskeleton and extracellular matrix molecules as well as in the activation of a variety of intracellular signalization processes in cooperation with growth factor receptors.Furthermore,recent studies show that distinct forms of α6 and β4 subunits are expressed in the human intestine and,more importantly,recent work provides experimental evidence that various forms of α6β4 can differentially regulate intestinal epithelial cell functions under both normal and pathological conditions.For instance,it has been discovered that colorectal cancer cells express a hybrid form of α6β4 that is never seen in normal cells.Although further work is needed,integrin α6β4 is emerging as a key regulator of intestinal functions in both intestinal health and disease.

异莲心碱通过PI3K/Akt/mTOR信号通路影响结肠癌SW480细胞增殖、凋亡和自噬

目的:探讨异莲心碱(Iso)通过PI3K/Akt/mTOR信号通路对结肠癌SW480细胞增殖、凋亡和自噬的影响。方法:用10、20和40μmol/L的Iso处理结肠癌SW480细胞,CCK-8法、流式细胞术和WB法分别检测Iso对细胞增殖活力、凋亡和自噬相关蛋白LC3Ⅰ、LC3Ⅱ、p62表达的影响。然后,用20μmol/L的Iso和25μmol/L的PI3K激活剂740 Y-P分别处理SW480细胞,将细胞分为对照组、740 Y-P组、Iso组和Iso+740 Y-P组,流式细胞术、WB法检测Iso和740 Y-P对各组细胞凋亡及细胞中LC3Ⅰ、LC3Ⅱ、p62、PI3K、p-PI3K、mTOR和p-mTOR蛋白表达的影响。结果:10、20和40μmol/L的Iso处理后,SW480细胞增殖活力均显著下降(均P<0.05),细胞凋亡率均显著升高(均P<0.05),LC3Ⅱ/LC3Ⅰ表达均显著上调(均P<0.05),p26蛋白表达显著下调(P<0.05)。Iso和740 Y-P处理后,与对照组相比,740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达均显著下降(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著升高(均P<0.05);Iso组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达升高(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著下降(均P<0.05);与740 Y-P组相比,Iso+740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达升高(P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著下降(均P<0.05);与Iso组相比,Iso+740 Y-P组细胞凋亡率、LC3Ⅱ/LC3Ⅰ表达下降(均P<0.05),p26、p-PI3K/PI3K和p-mTOR/mTOR表达均显著升高(均P<0.05)。结论:Iso通过抑制PI3K/Akt/mTOR信号通路抑制结肠癌SW480细胞增殖并诱导细胞凋亡和自噬。

奥氮平对乳腺癌相关巨噬细胞增殖与分化功能的影响

目的探讨奥氮平对乳腺癌细胞共培养体系中人急性髓系白血病细胞系THP-1细胞增殖和分化功能的影响。方法体外培养THP-1细胞、人乳腺癌MDA-MB-231细胞,建立THP-1与MDA-MB-231共培养组,用CCK-8法检测奥氮平处理后的THP-1单独组、共培养组中THP-1的活性变化;用qRT-PCR法检测奥氮平对佛波酯诱导的THP-1单独组、共培养组中THP-1表面分子CD68、CD80、CD163的表达,以及炎症因子IL-1β、IL-6、IL-12、TGF-α、TGF-β和IL-10的表达,同时进一步检测巨噬细胞极化相关miRNA分子miRNA9、miRNA155和miRNA34a的表达。结果奥氮平能抑制THP-1单独组和共培养组中THP-1的增殖活性(P<0.05);在佛波酯诱导THP-1向巨噬细胞分化的研究中,奥氮平能够显著提高THP-1单独组CD68分子的表达(P<0.05),而在共培养组中CD68、CD80、CD163的表达则均显著升高(P<0.05)。在THP-1单独组中,奥氮平仅能够促进IL-12的表达;在共培养组中,奥氮平使CCL2、CXCL10、IL-1β、IL-10、IL-6、IL-12、TNF-α表达水平显著升高(P<0.05)。在巨噬细胞分化相关miRNA的研究中,在奥氮平作用下,THP-1单独组中miR155和Let7c的表达显著升高(P<0.05),miR146、miR9和miR34a的表达变化无差异;在与MDA-MB-231共培养条件下,miR34a和Let7c显著升高(P<0.05)。结论在肿瘤细胞存在的情况下,奥氮平可参与肿瘤相关巨噬细胞表面分子、炎症因子以及巨噬细胞极化相关miRNA分子表达的调节,这为进一步利用奥氮平治疗乳腺癌提供了新思路。