Ca2+/calmodulin-dependent protein kinase II regulates colon cancer proliferation and migration via ERK1/2 and p38 pathways

AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immunochemistry. Transcriptional and posttranscriptional levels of Ca MKⅡin tissue samples and MMP2,MMP9 and TIMP-1 expression in the human colon cancer cell line HCT116 were assessed by q RTPCR and western blot. Cell proliferation was detected with the MTT assay. Cancer cell migration and invasion were investigated with the Transwell culture system and woundhealing assay.RESULTS We first demonstrated that CaMK Ⅱ was ove rexpressed in human colon cancers and was associated with cancer differentiation. In the human colon cancer cell line HCT116,the Ca MKII-specific inhibitor KN93,but not its inactive analogue KN92,decreased cancer cell proliferation. Furthermore,KN93 also significantly prohibited HCT116 cell migration and invasion. The specific inhibition of ERK1/2 or p38 decreased the proliferation and migration of colon cancer cells.CONCLUSION Our findings highlight Ca MKⅡ as a potential critical mediator in human colon tumor development and metastasis.

Stage migration vs immunology: The lymph node count story in colon cancer

Lymph node staging is of crucial importance for the therapy stratification and prognosis estimation in colon cancer. Beside the detection of metastases,the number of harvested lymph nodes itself has prognostic relevance in stage Ⅱ/Ⅲ cancers. A stage migration effect caused by missed lymph node metastases has been postulated as most likely explanation for that. In order to avoid false negative node staging reporting of at least 12 lymph nodes is recommended. However,this threshold is met only in a minority of cases in daily practice. Due to quality initiatives the situation has improved in the past. This,however,had no influence on staging in several studies. While the numbers of evaluated lymph nodes increased continuously during the last decades the rate of node positive cases remained relatively constant. This fact together with other indications raised doubts that understaging is indeed the correct explanation for the prognostic impact of lymph node harvest. Several authors assume that immune response could play a major role in this context influencing both the lymph node detectability and the tumor's behavior. Further studies addressing this issue are need. Based on the findings the recommendations concerning minimal lymph node numbers and adjuvant chemotherapy should be reconsidered.

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.

Luteolin inhibits the colon cancer HT-29 cell proliferation, migration and epithelial-mesenchymal transition: an experimental study

Objective: To study the regulating effect of luteolin on colon cancer HT-29 cell proliferation, migration and epithelial-mesenchymal transition. Methods: Colon cancer HT-29 cells were cultured and randomly divided into two groups, control group were treated with serum-free medium without drugs and LUT group were treated with serum-free medium containing luteolin. After 24 h of treatment, cells were collected to extract RNA, and then fluorescent quantitative PCR method was used to determine the mRNA expression of proliferation genes, migration genes and epithelial-mesenchymal transition genes. Results: After 24 h of luteolin treatment, Lrig1, TSPYL5, Bim, SOX15 and DLC1 mRNA expression in LUT group were significantly higher than those in control group while RPS15a, Bad, TRPV5, TRPV6, PLD2, IBP, SphK1, FAK, Vimentin and N-cadherin mRNA expression were significantly lower than those in control group. Conclusion: Luteolin has inhibiting effect on colon cancer HT-29 cell proliferation, migration and epithelial-mesenchymal transition.

Circular RNA PIP5K1A promotes colon cancer development through inhibiting mi R-1273a

BACKGROUND Circular RNAs (circRNAs) are considered to be highly stable due to the closed structure, which are predominately correlated with the development and progression of a wide variety of cancers. Colon cancer is one of the most common malignancies worldwide. A recent study demonstrated the upregulated expression of circPIP5K1A in non-small cell lung cancer. However, few studies have investigated the relationship between circ_0014130 level and colon cancer. Therefore, elucidating the underlying mechanisms of circPIP5K1A’s role may help with the identification of novel diagnostic and therapeutic targets for colon cancer. AIM To investigate the status of circPIP5K1A in colon cancers and its effects on the modulation of cancer development. METHODS The expression level of circPIP5K1A in tissue and serum samples from colon cancer patients, as well as human colonic cancer cell lines was detected by realtime quantitative reverse transcription-polymerase chain reaction. Following the transfection of specifically synthesized small interfering RNA (siRNA) into colon cell lines, we used Hoechst staining assay to measure the ratio of cell death in the absence of circPIP5K1A. Moreover, we also used the Transwell assay to assess the migratory function of colon cells overexpressing circPIP5K1A. Additionally, we employed a series of bioinformatics prediction programs to predict the potential of circPIP5K1A-targeted miRNAs and mRNAs. The miR-1273a vector was constructed, and then transfected with or without circPIP5K1A vector into colon cancer cells. Afterwards, the expression of activator protein 1 (AP-1), interferon regulating factor 4 (IRF-4), caudal type homeobox 2 (CDX-2), and zinc finger of the cerebellum 1 (Zic-1) was detected by western blotting. RESULTS CircPIP5K1A was significantly upregulated in colon cancer tissue relative to their adjacent normal tissues. Knockdown of circPIP5K1A in colon cancer cells impaired cell viability and suppressed cell invasion and migration, while enforced expression of circPIP5K1A exhibited the opposite effects on cell migration. Bioinformatics prediction program predicted that the association of circPIP5K1A with miR-1273a, as well as AP-1, IRF-4, CDX-2, and Zic-1. Subsequent studies showed that overexpression of circPIP5K1A augmented the expression of AP-1 but attenuated the expression of IRF-4, CDX-2, and Zic-1. Reciprocally, overexpression of miR-1273a abrogated the oncogenic function of circPIP5K1A in colon cancers. CONCLUSION Overall, our data demonstrate the oncogenic role of circPIP5K1A-miR-1273a axis in regulation of colon cancer development, which provides a novel insights into colon cancer pathogenesis.

Effects of lysophosphatidic acid on human colon cancer cells and its mechanisms of action

AIM： To study the effects of lysophosphatidic acid （LPA） on proliferation, adhesion, migration, and apoptosis in the human colon cancer cell line, SW480, and its mechanisms of action. METHODS： Methyl tetrazolium assay was used to assess cell proliferation. Flow cytometry was employed to detect cell apoptosis. Cell migration was measured by using a Boyden transweU migration chamber. Cell adhesion assay was performed in 96-well plates according to protocol. RESULTS： LPA significantly stimulated SW480 cell proliferation in a dose-dependent and timeependent manner compared with the control group （P 〈 0.05） while the mitogen-activated protein kinase （MAPK） inhibitor, PD98059, significantly blocked the LPA stimulation effect on proliferation. LPA also significantly stimulated adhesion and migration of SW480 cells in a dosedependent manner （P 〈 0.05）. Rho kinase inhibitor, Y-27632, significantly inhibited the upegulatory effect of LPA on adhesion and migration （P 〈 0.05）. LPA significantly protected cells from apoptosis induced by the chemotherapeutic drugs, cisplatin and 5-FU （P 〈 0.05）, but the phosphoinositide 3-kinase （PI3K） inhibitor, LY294002, significantly blocked the protective effect of LPA on apoptosis. CONCLUSION： LPA stimulated proliferation, adhesion,migration of 5W480 cells, and protected from apoptosis. The Ras/Raf-MAPK, G12/13-Rho-RhoA and PI3K- AKT/PKB signal pathways may be involved.

Outcomes of colon self-expandable metal stents for malignant vs benign indications at a tertiary care center and review of literature

BACKGROUND Endoscopic placement of a self-expandable metal stent(SEMS)is a minimally invasive treatment for use in malignant and benign colonic obstruction.However,their widespread use is still limited with a nationwide analysis showing only 5.4%of patients with colon obstruction undergoing stent placement.This underutilization could be due to perceived increase risk of complications with stent placement.AIM To review long-and short-term clinical success of SEMS use for colonic obstruction at our center.METHODS We retrospectively reviewed all the patients who underwent colonic SEMS placement over aeighteen year period (August 2004 through August 2022) at our academic center. Demographicsincluding age, gender, indication (malignant and benign), technical success, clinical success,complications (perforation, stent migration), mortality, and outcomes were recorded.RESULTSSixty three patients underwent colon SEMS over an 18-year period. Fifty-five cases were formalignant indications, 8 were for benign conditions. The benign strictures included diverticulardisease stricturing (n = 4), fistula closure (n = 2), extrinsic fibroid compression (n = 1), and ischemicstricture (n = 1). Forty-three of the malignant cases were due to intrinsic obstruction from primaryor recurrent colon cancer;12 were from extrinsic compression. Fifty-four strictures occurred on theleft side, 3 occurred on the right and the rest in transverse colon. The total malignant case (n = 55)procedural success rate was 95% vs 100% for benign cases (P = 1.0, NS). Overall complication ratewas significantly higher for benign group: Four complications were observed in the malignantgroup (stent migration, restenosis) vs 2 of 8 (25%) for benign obstruction (1-perforation, 1-stentmigration) (P = 0.02). When stratifying complications of perforation and stent migration there wasno significant difference between the two groups (P = 0.14, NS).CONCLUSIONColon SEMS remains a worthwhile option for colonic obstruction related to malignancy and has ahigh procedural and clinical success rate. Benign indications for SEMS placement appear to havesimilar success to malignant. While there appears to be a higher overall complication rate inbenign cases, our study is limited by sample size. When evaluating for perforation alone theredoes not appear to be any significant difference between the two groups. SEMS placement may bea practical option for indications other that malignant obstruction. Interventional endoscopistsshould be aware and discuss the risk for complications in setting of benign conditions. Indicationsin these cases should be discussed in a multi-disciplinary fashion with colorectal surgery.

NKD1促进结肠癌细胞迁移的机制研究及临床意义

目的:探讨NKD1在结肠癌组织中的表达及其与结肠癌患者临床病理特征因素间的相关性,以及研究其在结肠癌细胞迁移中的作用,并明确其调控上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白ZEB1的具体机制,为转移性结肠癌提供新的治疗靶点。方法:使用生物信息学初步分析NKD1在结肠癌细胞中的表达情况及其与临床病理特征因素间的相关性。通过收集98对来自常州市武进人民医院的结肠癌患者组织样本及相关临床信息验证生物信息学结果,绘制生存曲线。划痕实验、Transwell实验证明NKD1促进结肠癌细胞迁移。qPCR、western blot实验验证NKD1在蛋白水平调控ZEB1。Western blot、免疫沉淀等实验深入研究NKD1调控ZEB1的具体机制。结果:NKD1在结肠癌组织中高表达,其表达水平与分化程度、肿瘤T分期及是否远处转移等临床病理特征因素具有相关性;NKD1高表达与结肠癌患者的不良预后相关;过表达NKD1后促进结肠癌细胞的迁移,敲除NKD1后则结果相反;NKD1在蛋白水平调控ZEB1,并能够维持ZEB1的蛋白稳定性;高表达NKD1通过调控ZEB1与自噬相关蛋白LC3的结合抑制ZEB1的自噬降解。结论:NKD1与结肠癌患者的不良预后相关,并且可以通过调控ZEB1与自噬相关蛋白LC3的结合抑制ZEB1的自噬降解,从而促进结肠癌细胞的迁移。

地榆皂苷Ⅱ抑制结肠癌细胞增殖、迁移、侵袭和诱导凋亡的体外实验研究

目的探讨地榆皂苷Ⅱ对结肠癌细胞HT-29增殖、迁移、侵袭和凋亡的作用并探讨其作用机制。方法采用CCK-8法检测地榆皂苷Ⅱ对细胞增殖的影响,采用划痕试验检测地榆皂苷Ⅱ对细胞迁移能力的影响,采用Transwell小室实验检测地榆皂苷Ⅱ对细胞侵袭能力的影响,采用流式细胞术测定地榆皂苷Ⅱ对细胞凋亡的影响,分别采用实时定量反转录聚合酶链反应(qRT-PCR)和Western blot法测定地榆皂苷Ⅱ对细胞中蛋白激酶B(AKT)/磷脂酰肌醇-3-激酶(PI3K)信号通路mRNA和蛋白表达的影响。结果地榆皂苷Ⅱ(0、1、5、10、20、40、60和80μmol/mL)可剂量依赖性抑制结肠癌细胞HT-29的增殖;地榆皂苷Ⅱ(5、10和20μmol/mL)可剂量依赖性抑制结肠癌细胞HT-29的迁移能力;地榆皂苷Ⅱ(5、10和20μmol/mL)可剂量依赖性抑制结肠癌细胞HT-29的侵袭能力;地榆皂苷Ⅱ(5、10和20μmol/mL)可剂量依赖性促进结肠癌细胞HT-29的凋亡;地榆皂苷Ⅱ(5、10和20μmol/mL)可剂量依赖性降低结肠癌细胞HT-29中AKT和PI3K mRNA的表达,增加Caspase-3和Caspase-9 mRNA的表达;地榆皂苷Ⅱ(5、10和20μmol/mL)可剂量依赖性地降低结肠癌细胞HT-29中磷酸化蛋白激酶B(p-AKT)和磷酸化磷脂酰肌醇-3-激酶(p-PI3K)蛋白的表达,增加Cleaved-Caspase-3和Cleaved-Caspase-9蛋白的表达,差异有统计学意义(P<0.05)。结论地榆皂苷Ⅱ具有抑制结肠癌细胞HT-29的增殖、迁移、侵袭,促进细胞凋亡作用,可能与其促进AKT和PI3K蛋白磷酸化、Caspase-3和Caspase-9蛋白活化而调节AKT/PI3K信号通路有关。

SETD5介导AKT1磷酸化调控结肠癌细胞的迁移和5-FU敏感性

目的:探讨含SET结构域蛋白5(SETD5)对结肠癌细胞增殖、迁移和对5-氟尿嘧啶(5-FU)药物敏感性的影响及机制。方法:常规培养结肠癌细胞,用Lipofectamine 2000将siSETD5-NC、si-SETD5-1~3质粒转染至HT-29细胞中,将其分为对照组(未处理)、si-SETD5-NC组、si-SETD5组和si-SETD5+SC79组,si-SETD5+SC79组HT-29细胞转染质粒的同时用10µmol/L SC79处理。qPCR法检测NCM460、HT-29和LoVo细胞中SETD5 mRNA表达,流式细胞术、细胞划痕法、WB法和CCK-8法分别检测各组HT-29细胞的凋亡情况、迁移能力、相关蛋白的表达,以及对5-FU的敏感性。结果:SETD5 mRNA在HT-29、LoVo细胞中均呈高表达(均P<0.01)。在HT-29细胞中成功地敲减了SETD5 mRNA(P<0.01)。敲减SETD5 mRNA可明显抑制HT-29细胞的增殖活性(P<0.01)、迁移能力(P<0.01)、相关蛋白(SETD5、p-PI3K、p-AKT1、p-mTOR蛋白)的表达(均P<0.01)、促进细胞凋亡(P<0.01),且提高其对5-FU的敏感性(P<0.01),这些作用均可被AKT激活剂SC79部分阻挡(P<0.05或P<0.01)。结论:SETD5在HT-29、LoVo细胞中高表达,SETD5通过PI3K/AKT1通路促进结肠癌HT-29细胞的增殖、迁移,且降低其对5-FU的敏感性,SETD5是结肠癌临床诊断、治疗的潜在靶点。

STAT3抑制剂stattic对小鼠结肠癌CT26细胞增殖和凋亡的影响

目的探讨信号转导和转录激活因子3(STAT3)抑制剂盐酸萘替芬(stattic)对小鼠结肠癌CT26细胞增殖和凋亡的影响和作用机制。方法采用0μmol/L、1μmol/L、5μmol/L、10μmol/L stattic溶液处理小鼠结肠癌CT26细胞,通过CCK-8实验、细胞克隆形成实验、细胞划痕实验、Transwell侵袭实验以及流式细胞术检测细胞活力、增殖、迁移、侵袭、周期和凋亡情况;利用Western blot法检测stattic对小鼠结肠癌细胞磷酸化STAT3(p-STAT3)表达的影响;通过实时荧光定量多聚核苷酶链式反应(RT-qPCR)检测stattic作用后CT26细胞B淋巴细胞瘤-2(Bcl-2)和人跨膜受体蛋白Notch-1(Notch-1)的表达。结果与0μmol/L组相比,stattic溶液组CT26细胞的活力及增殖能力降低(P<0.001)、迁移率和侵袭率降低(P<0.001),细胞凋亡率随浓度增加而增加(P<0.0001);stattic能将CT26细胞周期阻断于G1期,进而阻止CT26细胞的增殖;Western blot结果显示stattic抑制CT26细胞p-STAT3的表达(P<0.05);RT-qPCR检测结果表明stattic下调CT26细胞Bcl-2和Notch1的表达(P<0.05)。结论stattic通过阻断STAT3信号,抑制p-STAT3蛋白的表达,下调下游抗凋亡分子Bcl-2和Notch1信号分子的表达从而抑制CT26细胞增殖促进细胞凋亡。

SERPINE1对结肠癌细胞增殖、迁移、侵袭及凋亡的影响

目的探究丝氨酸蛋白酶抑制剂1(serine proteinase inhibitor family E member 1,SERPINE1)在结肠癌中的表达及对结肠癌细胞恶性生物学行为的影响。方法通过starBase和TISIDB数据库分析471个结肠癌及41个正常结肠组织中SERPINE1的表达及其与临床分期、预后生存的关系;收集联勤保障部队第九四〇医院5例结肠癌患者癌组织与对应的癌旁组织标本,应用RT-qPCR和Western blot验证SERPINE1在结肠癌中的表达。构建SERPINE1过表达与干扰表达的慢病毒载体,进行结肠癌RKO和LoVo细胞株转染,通过RT-qPCR和Western blot评估SERPINE1过表达和干扰表达的效果,通过细胞计数试剂盒(cell counting kit-8,CCK-8)、克隆形成实验、Transwell实验和Hoechst凋亡细胞核染色评估细胞的增殖、迁移与侵袭能力和细胞的凋亡情况;应用Western blot检测SERPINE1对磷脂酰激酶3(phosphatidyl-inositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路的影响。结果数据库分析发现SERPINE1在结肠癌中高表达,且与患者的肿瘤临床分期正相关,与预后总生存期负相关(P<0.05)。转染过表达的慢病毒载体后细胞的增殖、迁移与侵袭能力增强,而细胞的凋亡减弱;反之,在转染干扰表达的慢病毒载体后细胞的增殖、迁移与侵袭能力减弱,而细胞的凋亡增强(P<0.05)。通过Western blot实验发现转染SERPINE1过表达的慢病毒载体后细胞PI3K和AKT表达无明显变化,而磷酸化PI3K(p-PI3K)和磷酸化AKT(p-AKT)表达增加,反之转染SERPINE1干扰表达的慢病毒载体细胞PI3K和AKT表达无明显变化,而p-PI3K和p-AKT表达减少,灰度值定量分析p-PI3K/PI3K和p-AKT/AKT比值差异有统计学意义(P<0.05)。结论SERPINE1在结肠癌中高表达,与患者临床分期及预后相关;SERPINE1过表达可能通过激活PI3K/AKT信号通路促进结肠癌细胞的增殖、迁移与侵袭,抑制其凋亡。

LINC00638在结肠癌中的表达及对结肠癌细胞功能的研究

目的探讨LINC00638在结肠癌中的表达及其临床意义,并进一步探究LINC00638对结肠癌细胞生物学功能的影响。方法通过癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库初步比较LINC00638在正常结肠组织和结肠癌组织中的表达差异,探究其与患者临床病理特征的关系。通过Kaplan-Meier生存分析进一步探究LINC00638表达对结肠癌患者预后的影响。通过GEPIA数据库和高通量基因表达数据库(Gene Expression Omnibus,GEO)进一步验证LINC00638表达对结肠癌患者预后的影响。采用单因素和多因素COX回归分析探究LINC00638、临床病理相关因素对结肠癌患者预后的影响。应用小干扰RNA(siRNA)沉默HCT116细胞和DLD1细胞中LINC00638的表达,通过CCK-8实验、集落形成实验、划痕实验、Transwell实验探究LINC00638对结肠癌细胞增殖、迁移和侵袭能力的影响。结果LINC00638在结肠癌组织中表达水平明显升高,并且其表达量与结肠癌患者的淋巴结转移和TNM分期密切相关。Kaplan-Meier生存曲线分析结果显示,LINC00638表达水平越高,结肠癌患者预后越差(P<0.05)。COX回归分析结果显示,LINC00638、年龄和TNM分期可以作为结肠癌患者的独立预后因素。细胞功能实验结果表明,与对照组比较,敲低组细胞的增殖、迁移和侵袭能力显著降低(P<0.05)。结论LINC00638在结肠癌中高表达,并且其表达水平越高,结肠癌患者预后越差。敲低LINC00638后可以抑制结肠癌细胞的增殖、迁移和侵袭能力。

MiR-152-3p调控DNA甲基转移酶1对结肠癌增殖、迁移、侵袭和凋亡的影响

目的:探究miR-152-3p、DNA甲基转移酶1(DNA methyltransferases,DNMT1)在结肠癌细胞中的调控机制。方法:qRT-PCR检测miR-152-3p和DNMT1的表达;细胞增殖实验、克隆形成实验、划痕愈合实验以及Transwell实验检测各处理组癌细胞的增殖、迁移和侵袭能力;双荧光素酶实验检测miR-152-3p与DNMT1的靶向关系;Western blot实验检测DNMT1蛋白表达;流式细胞术实验检测细胞凋亡率。结果:结肠癌细胞中miR-152-3p显著低表达,DNMT1显著高表达;过表达miR-152-3p会抑制结肠癌细胞增殖、迁移和侵袭;miR-152-3p能靶向抑制DNMT1的表达,从而抑制结肠癌细胞的增殖、迁移和侵袭;过表达miR-152-3p可以靶向DNMT1并促进细胞凋亡。结论:miR-152-3p通过靶向下调DNMT1的表达,从而抑制结肠癌细胞的发生发展。

miR-141-3p靶向MYT1L促进结肠癌细胞的恶性进展

目的:探究微小RNA-141-3p(miR-141-3p)通过靶向作用髓磷脂转录因子1(MYT1L)蛋白对结肠癌细胞迁移、增殖、凋亡的影响。方法:在结肠癌HCT8细胞中转染miR-141-3p模拟物(miR-141-3p mimic),qRT-PCR检测miR-141-3p的mRNA表达水平,流式细胞术检测细胞凋亡,Transwell法检测侵袭和迁移,Western blot检测MYT1L的蛋白表达水平。生物信息学软件预测MYT1L可能是miR-141-3p的靶基因后用荧光素酶报告基因鉴定。结果:miR-141-3p在结肠癌组织细胞中显著上调且对患者预后有显著影响,在转染了miR-141-3p的HCT8细胞中,细胞的增殖、迁移能力均显著提高,而细胞凋亡显著下降。双荧光素酶报告基因的结果显示miR-141-3p与MYT1L基因可以靶向结合。共转染MYT1L过表达载体和miR-141-3p mimic的细胞中,相对于只转染了MYT1L过表达载体的细胞来说,细胞表型恶化程度显著升高。结论:miR-141-3p通过靶向调控MYT1L的表达促进结肠癌细胞表型的恶化。

G protein-coupled estrogen receptor in colon function, immune regulation and carcinogenesis

Estrogens play important roles in the development and progression of multiple tumor types.Accumulating evidence points to the significance of estrogen action not only in tumors of hormonally regulated tissues such as the breast,endometrium and ovary,but also in the development of colorectal cancer(CRC).The effects of estrogens in physiological and pathophysiological conditions are mediated by the nuclear estrogen receptorsαandβ,as well as the membranebound G protein-coupled estrogen receptor(GPER).The roles of GPER in CRC development and progression,however,remain poorly understood.Studies on the functions of GPER in the colon have shown that this estrogen receptor regulates colonic motility as well as immune responses in CRC-associated diseases,such as Crohn’s disease and ulcerative colitis.GPER is also involved in cell cycle regulation,endoplasmic reticulum stress,proliferation,apoptosis,vascularization,cell migration,and the regulation of fatty acid and estrogen metabolism in CRC cells.Thus,multiple lines of evidence suggest that GPER may play an important role in colorectal carcinogenesis.In this review,we present the current state of knowledge regarding the contribution of GPER to colon function and CRC.

Verticillin A inhibits colon cancer cell migration and invasion by targeting c-Met

Verticillin A is a diketopiperazine compound which was previously isolated from Amanita flavorubescens Aik(containing parasitic fungi Hypomyces hyalines(Schw.)Tul.).Here,we initially found,by wound healing assay and Tran swell assay in vitro,that verticilli n A possesses an inhibitory effect agai nst the migrati on and in vasion of the human colon cancer cell.Subsequently,c-mesenchymal,epithelial transition factor(c-Met)was identified as a molecular target of verticillin A by screening key genes related to cell migration.Verticillin A-mediated c-Met suppress!on is at the transcriptio nal level.Further study dem on strated that verticilli n A suppressed c-MET phosphorylation and decreased c-MET protein level.In addition,verticillin A inhibited the phosphorylation of c-MET downstream molecules including rat sarcoma(Ras)-associated factor(Raf),extracellular signal-regulated kinase(ERK),and protein kinase B(AKT).Overexpression of Erk partially reversed the verticillin A-mediated anti-metastasis action in the human colon cancer cell.More importantly,verticillin A also inhibited cancer cell metastasis in vivo.Thus,verticillin A can significantly inhibit the migration and invasion of colon cancer cells by targeting c-Met and inhibiting Ras/Raf/mitogen-activated extracellular signal-regulated kinase(MEK)/ERK signaling pathways.Therefore,we determined that verticillin A is a natural compound that can be further developed as an anti-metastatic drug in human cancers.

MiR-152-3p靶向调控KLF4促进结肠癌细胞增殖、迁移和侵袭

目的:探讨微小RNA-152-3p(miR-152-3p)在结肠癌(CC)中的表达情况及靶向Krüppel样因子4(KLF4)对CC细胞的增殖、迁移和侵袭的影响。方法:通过starBase及TCGA数据库分析预测miR-152-3p与KLF4之间的靶向关系,以及各自在正常组织和CC组织中的表达情况差异;双荧光素酶和RIP实验验证miR-152-3p与KLF4之间的靶向关系;RT-q PCR检测miR-152-3p与KLF4在CC细胞系中的mRNA表达量;Western blot实验检测KLF4在CC细胞系中的蛋白表达水平;MTT实验检测细胞的增殖能力;划痕愈合实验检测细胞的迁移能力;Transwell实验检测细胞的侵袭能力;细胞周期实验和凋亡实验检测细胞的周期分布情况和凋亡百分率。结果:miR-152-3p在CC组织和细胞系中高表达(P<0.001),而KLF4低表达(P<0.01),且miR-152-3p能够靶向下调KLF4的表达(P<0.01)。miR-152-3p过表达能增高CC细胞系的增殖、迁移和侵袭能力,降低细胞周期在G0/G1期的分布,并抑制其凋亡(P<0.05);KLF4过表达可以抑制miR-152-3p对CC细胞增殖、迁移和侵袭能力的促进作用(P<0.05),使G0/G1期细胞比例增加(P<0.05),并促进细胞凋亡(P<0.05)。结论:miR-152-3p通过靶向下调KLF4的表达促进CC细胞的增殖、迁移和侵袭。

双歧杆菌对结肠癌细胞DLD-1增殖、侵袭及迁移的影响

目的通过体外研究初步评估双歧杆菌对结肠癌细胞增殖、侵袭及迁移的影响。方法以人源结肠癌细胞DLD-1为研究对象,通过CCK8、划痕及Transwell实验探究双歧杆菌对结肠癌细胞DLD-1增殖、侵袭及迁移的影响。结果CCK8实验表明,相比于对照组,双歧杆菌处理后的结肠癌细胞DLD-1的增殖能力被显著抑制(P<0.01);划痕实验结果表明,相比于对照组,双歧杆菌可以显著抑制结肠癌细胞DLD-1的迁移能力(P<0.01);Transwell实验表明,相比于对照组,双歧杆菌可以显著抑制结肠癌细胞DLD-1的侵袭能力(P<0.01)。结论双歧杆菌可以减弱结肠癌细胞DLD-1增殖、侵袭及迁移能力,本研究为临床结肠癌治疗提供新的思路。

环状RNA_0053063通过ATG13依赖的途径调控结肠癌细胞的迁移和侵袭能力

目的:探讨环状RNA_0053063(circ_0053063)在结肠癌细胞中的表达及其对结肠癌细胞迁移和侵袭能力的影响与调控机制。方法:通过qRT-PCR检测circ_0053063在正常肠上皮NCM460细胞和肠癌LoVo、HCT116、SW480、CACO-2和HT29细胞中的表达水平。构建稳定过表达circ_0053063的肠癌LoVo细胞并应用瞬时转染shATG13的方法干扰ATG13的表达再应用脂多糖预处理肠癌LoVo细胞,通过细胞划痕实验和Transwell实验检测各组LoVo肠癌细胞的迁移和侵袭能力。结果:circ_0053063在正常肠上皮NCM460细胞呈高表达,而在肠癌LoVo、HCT116、SW480、CACO-2和HT29细胞中呈低表达。肠癌LoVo细胞,稳定过表达circ_0053063可显著促进肠癌LoVo细胞的迁移和侵袭能力。瞬时干扰ATG13后抑制过表达circ_0053063导致的肠癌LoVo细胞的迁移和侵袭能力。结论:circ_0053063通过ATG13依赖的途径调控肠癌细胞的迁移和侵袭能力。