人细胞色素P450 前mRNA的可变剪接研究进展

Human genes typically contain multiple introns, and in many cases the exons can be joined more than one way to generate multiple rnRNAs, encoding distinct protein isoforms. This process is called alternative splicing. The article summarized the human cytochrome P450 pre-mRNA alternative splicing and their regulatory mechanism and impacts on biological functions.     

剪接因子MBNL3对胰腺癌细胞恶性生物学行为的影响

目的 探讨MBNL3在胰腺癌细胞中的表达及对其恶性生物学行为的影响.方法 收集自2017年至2020年北京协和医院50例胰腺癌石蜡组织样本,采用免疫组织化学法检测MBNL3蛋白表达.胰腺癌细胞经过MBNL3表达或敲低后,采用实时无标记细胞检测技术、平板集落形成技术和Transwell小室法验证MBNL3对细胞增殖、细胞...     

mRNA的差异剪接与神经系统发育

mRNA的选择性剪接可以使基因产生新的结构与功能,是基因表达调控的重要机制.神经系统中大部分的差异剪接基因与信号传导和调节有关,包括转录因子、受体、离子通道等,这些差异剪接影响细胞内基因的转录调控、信号传导、离子通道的动力学,从而对神经系统的发育、功能以及内环境的稳定性具有重要的调节作用.差异剪接的异常导致神经系统的疾病和肿瘤,因此可以作为治疗的靶标.     

Foxp3和PD1在结直肠癌组织中的表达及其临床意义

目的 检测叉头状转录因子3(Foxp3)和程序性死亡受体1(PD1)在结直肠癌组织中的表达,探讨两者表达与患者临床病理特征及预后的关系.方法 应用免疫组化(IHC)技术检测Foxp3及PD1在结直肠癌组织中和癌旁组织中的表达,Chi-square统计分析Foxp3及PD1表达强度的相关性及与患者临床病理特征,Kapla...     

PLIN3在肺腺癌组织和细胞中的表达及与预后的关系

目的:研究围脂滴蛋白3(PLIN3)在肺腺癌组织和细胞中的表达情况,分析其与患者预后的关系,以及其对肺腺癌细胞侵袭能力的影响。方法:使用HPA数据库预测与肺癌不良预后相关的基因,选中PLIN3为研究对象,并分析PLIN3表达与肺癌及肺腺癌患者生存率的相关性;运用GEPIA数据库验证肺腺癌中PLIN3的表达差异及其与肺腺癌患者生存率的相关性;运用Ualcan数据库进一步分析PLIN3在肺腺癌中的表达情况;Western blot实验检测PLIN3在肺腺癌细胞中的表达水平;构建敲减PLIN3表达的质粒siPLIN3,采用Transwell实验检测转染后A549细胞的侵袭能力。结果:PLIN3与肺腺癌患者生存率显著相关,其在肺腺癌组织中高表达,且与肿瘤分级、淋巴结转移和分期密切相关,与患者性别无关。PLIN3在肺腺癌细胞中高表达,敲减PLIN3表达后,敲减组细胞穿过Transwell小室的数量明显低于未敲减组。结论:PLIN3在肺腺癌组织中高表达,其表达水平与肺腺癌患者的预后相关;且PLIN3可以促进肺腺癌细胞的侵袭。     

circPSMC3在结直肠癌组织中的表达及对CRC细胞增殖和转移的影响

目的 研究环状RNAPSMC3(circPSMC3)在结直肠癌(CRC)组织中的表达水平及临床意义,探讨circPSMC3对CRC细胞增殖和转移的作用及作用机制.方法 采用荧光定量PCR(qRT-PCR)检测circPSMC3在CRC和癌旁组织中的表达水平,分析circPSMC3的表达与CRC患者临床病理参数的关系.采...     

STAT3与结直肠癌的研究进展

STAT3是信号转导和转录因子家族（STAT）的重要成员,经细胞外细胞因子、生长因子等多种刺激信号激活,作用于细胞核内特异的DNA片段,调控靶基因的转录。近年来研究表明,STAT3在结直肠癌的发生、发展中起重要的作用。通过调控STAT3可能成为结直肠癌分子治疗的新靶点。     

WNT信号抑制因子DKK3基因 对结直肠癌细胞系HCT116迁移与侵袭的影响

目的分析DKK3在结直肠癌组织中的表达及对患者预后的影响,同时探索DKK3对结直肠癌细胞增殖、迁移以及侵袭能力的影响,并对机制进行一定的探索。方法在线数据库分析DKK3在结直肠癌组织中的表达及其对患者预后的影响。用RNA干扰技术,下调DKK3;用MTT检测细胞增殖,同时运用细胞小室迁移实验检测细胞迁移及侵袭能力,并通过RT-qPCR技术以及Western blot技术检测迁移相关蛋白表达。结果与正常组织相比,DKK3在结直肠癌组织中的表达显著上调并会导致患者整体预后情况变差(P0.05),干涉DKK3后细胞增殖速率明显减缓(P0.05),并且细胞迁移以及侵袭能力减弱(P0.01)。MMP2、 MMP7以及MMP9与DKK3的表达呈正相关关系(P0.001)。在敲低DKK3后,MMP2、 MMP7、 MMP9以及p-ERK的表达均显著下调(P0.01)。结论 DKK3在结直肠癌细胞系HCT116中表达上调,可能通过MAPK/ERK信号通路促进细胞增殖以及迁移能力。     

局部浸润Foxp3+调节性T细胞在结直肠癌中的研究进展

肿瘤微环境中的免疫状态对肿瘤患者的预后意义重大.调节性T细胞(Tregs)是造成肿瘤免疫逃逸和肿瘤免疫治疗失败的关键因素.Foxp3是Tregs细胞重要的胞内信号标记,并在Tregs细胞的发育和功能中起关键作用.大多数肿瘤中局部浸润Tregs细胞的升高往往提示预后不良,但是在结直肠癌中Tregs细胞的预后意义各家报道仍有分歧.Tregs细胞的多态性包括表型、功能等,可能是造成这一矛盾的原因之一,但是缺乏一种更具特异性的标记或者标准化的检测手段也可能是造成这一现象的原因.     

miR-625-3p在结肠癌组织和细胞中的表达

目的:探讨微小RNA(mi R)-625-3p在结肠癌组织和细胞中的表达,了解其作用机制。方法:利用q RT-PCR分析结肠癌细胞株、癌组织和癌旁组织中mi R-625-3p的表达水平,探讨mi R-625-3p表达水平与结肠癌临床病理参数之间的关系;利用碘化丙啶染色和流式细胞术检测细胞凋亡和细胞周期变化;Western blot法检测mi R-625-3p对结肠癌细胞株凋亡相关蛋白的影响,初步了解其发挥影响的可能机制。结果:结肠癌组织mi R-625-3p的表达量比癌旁组织高(P0.05),mi R-625-3p的表达量与结肠癌浸润深度、TNM分期及有无远处转移关系密切(P0.05)。mi R-625-3p在结肠癌SW620细胞中的表达水平高于在SW480细胞中的表达。抑制mi R-625-3p的表达能显著抑制结肠癌细胞的周期(P0.05)和促进凋亡;转染mi R-625-3p后,Bcl-2的表达量增加(P0.05),Bax表达量没有显著变化。结论:mi R-625-3p能促进结肠癌细胞增殖,抑制结肠癌细胞凋亡,可能是结肠癌新的抗癌靶点。     

Regulation between two alternative splicing isoforms ZNF148FL and ZNF148ΔN, and their roles in the apoptosis and invasion of colorectal cancer

Objective

To investigate the effect of two alternative splicing isoforms of zinc finger protein (ZNF) 148 gene on the invasion and metastasis of human colorectal cancer (CRC) cells and their related mechanisms.

Epsin3在结直肠癌中的差异表达及生物信息学分析

目的 探讨介导内吞作用的衔接蛋白epsin 3（EPN3）在结直肠癌中的表达及意义,为深入研究EPN3的调控模式提供实验依据。 方法 分别运用GEPIA和GEDS数据库分析EPN3在结直肠癌组织和细胞中的表达情况,并通过SMART和cBioPortal数据库分析EPN3基因甲基化和拷贝数变异与其表达水平的关系;利用Metascape数据库对EPN3相关的共表达基因集进行GO富集和通路分析;运用BioPlex蛋白互作数据库分析EPN3在HCT116细胞中的蛋白作用网络。为了对EPN3进一步验证,我们收集13对结直肠癌癌旁组织和癌组织标本,用Real-time PCR检测EPN3 mRNA表达;并通过敲减EPN3观察其对肿瘤细胞增殖、集落形成和迁移能力的影响。 结果 GEPIA、GEDS、SMART和cBioPortal等数据库分析显示,EPN3在结直肠肿瘤组织中高表达(P<0.01)。其表达水平与甲基化和拷贝数变异相关。EPN3相关基因的富集结果显示主要与细胞黏附相关。EPN3与UBB、CCDC130、TNFAIP1、PHGDH、EPN2等构成的蛋白相对作用网络与蛋白泛素化有关。Real-time PCR结果显示,EPN3在癌组织中高表达(P<0.05)。通过沉默EPN3可以抑制HCT116和HT29细胞的增殖、集落形成和迁移能力。 结论 EPN3在结直肠癌组织中高表达,且与细胞黏附和蛋白泛素化等生物学过程有关,敲低EPN3可抑制结直肠癌细胞系HCT116和HT29的增殖、集落形成和迁移等过程。     

高尔基体磷蛋白3在结直肠癌组织中表达及其对结直肠癌细胞增殖、侵袭和迁移的影响

目的 分析高尔基体磷蛋白3（GOLPH3）在结直肠癌组织中的表达情况,及其对结直肠癌细胞增殖、侵袭和迁移的影响。方法 收集术前未经放、化疗治疗,经手术切除患者的结直肠癌组织及其相应的癌旁组织（40例）;采用免疫组织化学法检测组织中GOLPH3的表达,分析GOLPH3与结直肠癌临床病理特征的关系;构建稳定沉默GOLPH3的结直肠癌细胞株,并设立阴性对照组和空白对照组,Western blotting法检测3组细胞中GOLPH3蛋白的表达,MTT法检测3组细胞的增殖活性,Transwell实验分别检测3组细胞的侵袭和迁移能力。结果 GOLPH3在癌组织中主要为阳性表达,且阳性表达率（65.0%）显著高于癌旁组织（35.0%）（P＜0.05）;GOLPH3在TNM分期为Ⅲ～Ⅳ期、浸润深度≥2 cm、中低分化及有淋巴结转移的癌组织中的阳性表达率明显高于TNM分期Ⅰ～Ⅱ期、浸润深度＜2 cm、高分化及无淋巴结转移的癌组织（P＜0.05）。与阴性对照组和空白对照组比,基因沉默组GOLPH3蛋白表达量、细胞增殖活性、侵袭能力及迁移能力均明显降低（P＜0.05）。结论 GOLPH3在结直肠癌组织中高表达,其表达与肿瘤分期、侵袭深度、分化程度及淋巴结转移有关,抑制GOLPH3基因表达可下调GOLPH3蛋白表达,抑制结直肠癌细胞增殖、侵袭和迁移。     

Alternative pre-mRNA splicing regulation in cancer: pathways and programs unhinged

Alternative splicing of mRNA precursors is a nearly ubiquitous and extremely flexible point of gene control in humans. It provides cells with the opportunity to create protein isoforms of differing, even opposing, functions from a single gene. Cancer cells often take advantage of this flexibility to produce proteins that promote growth and survival. Many of the isoforms produced in this manner are developmentally regulated and are preferentially re-expressed in tumors. Emerging insights into this process indicate that pathways that are frequently deregulated in cancer often play important roles in promoting aberrant splicing, which in turn contributes to all aspects of tumor biology.     

结肠癌组织中Ski的表达及其与临床病理因素的关系

目的 研究Ski在结肠癌组织中的表达与临床病理因素的相关性.方法 采用免疫组化检测癌旁和结肠癌组织中Ski蛋白表达情况.结果 结肠癌组织中Ski蛋白表达阳性率[61.76％(21/34)]明显高于癌旁组织[38.24％(13/34)],差异具有统计学意义(P=0.02).Ski阳性表达与肿瘤的浸润深度(P=0.03)、有无淋巴结转移(P=0.005)和患者的预后有关(P=0.005),而与患者的性别、年龄、肿瘤分化程度无关(P＞0.05).结论 Ski在结肠癌组织中的表达明显增强,并且与肿瘤的浸润、转移和患者的预后有关.     

丙泊酚对结直肠癌细胞生物学行为的影响

目的:研究丙泊酚对结直肠癌细胞活力、侵袭和凋亡的影响,并探讨其作用机制。方法:10、25、50和100μmol/L的丙泊酚处理Lo Vo细胞72 h,或以100μmol/L的丙泊酚处理细胞12、24、48和72 h,CCK-8实验检测细胞活力;Transwell小室检测经100μmol/L丙泊酚处理72 h的细胞的侵袭能力;流式细胞术检测细胞周期及细胞凋亡率;Western blot检测基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、活化的胱天蛋白酶3(cleaved caspase-3)、Notch1和发状分裂相关增强子1(Hes1)的蛋白表达。结果:丙泊酚可抑制结直肠癌细胞活力;丙泊酚组细胞侵袭能力、S期细胞及MMP-2、MMP-9、Notch1和Hes1蛋白表达均显著低于对照组,而细胞凋亡率、G0/G1期细胞及cleaved caspase-3的蛋白水平均显著高于对照组(P0.01)。结论:丙泊酚可抑制结直肠癌Lo Vo细胞生长及侵袭能力,阻滞细胞周期,并诱导细胞凋亡,其机制与下调Notch1信号通路有关。     

Differential expression of hMLH1 in sporadic human colorectal cancer tumors and distant metastases

Somatic defects in the mismatch repair system constitute an important pathway in colorectal carcinogenesis. We have examined the expression of mismatch repair proteins in sporadic stage IV colorectal tumors and their derived metastases. Sporadic tumors were further examined for differences in expression between the tumor transition zone and the invasive front. Expression of hMSH2, hMLH1, and hPMS2 was screened immunohistochemically in 92 stage IV tumors and derived liver metastases. In cases with loss of mismatch repair protein expression, lymph node metastases were also examined. Clinicopathological parameters and Ki‐67 staining indexes were evaluated and compared. Four tumors displayed a complete loss of hMLH1/hPMS2 expression at the transition zone; however, three of these expressed both proteins at the invasive front and in liver and lymph node metastases. A further four were predominantly hMLH1/hPMS2 negative at the transition zone, but with distinct subclones of hMLH1/hPMS2‐expressing cells at the transition zone. All of these tumors expressed hMLH1/hPMS2 at the invasive front and in liver metastases, with three also expressing hMLH/hPMS2 in lymph node metastases. No significant difference in the proliferative index was observed for the hMLH1/hPMS2‐compromised group. In stage IV tumors re‐expression of hMLH1/hPMS2 occurred, leading to different patterns of expression within the primary tumor and between tumor and metastases. This may have functional importance for the chemosensitivity of metastases compared to the primary tumor.     

Differential expression of p63 isoforms in normal tissues and neoplastic cells

The p63 gene encodes at least six different proteins with homology to the tumour suppressor protein p53 and the related p53 family member p73. So far, there have been limited data concerning the expression patterns of individual p63 proteins, due to a lack of reagents that distinguish between the different isoforms. Three antibodies have been produced specifically directed against the two N-terminal isoforms (TAp63 and DeltaNp63) and the C-terminal region of the p63alpha proteins. TAp63 proteins are located suprabasally in stratified epithelia compared with the N-terminal truncated forms, which are more abundantly expressed in the basal cell layer, indicating a switch in expression of p63 isoforms during normal cellular differentiation. Analysis of squamous cell carcinomas shows DeltaNp63alpha to be the most widely expressed isoform, compatible with a role for this protein in promoting neoplastic cell growth in these tissues. DeltaNp63 protein expression is also restricted to basal cells in breast and prostate, whilst TAp63 isoforms are more widely expressed in these tissues as well as in tumours at these sites. TAp63, but not DeltaNp63 or p63alpha, is detected in normal colon and in colon carcinoma. TAp63 proteins are also expressed in the nuclei of a sub-population of lymphoid cells and in most malignant lymphomas, whereas DeltaNp63 proteins are not expressed. Taken together, a hitherto unrecognized regulation of p63 isoform expression in vivo has been uncovered, with different p63 proteins expressed during differentiation and in different cell types. The data indicate roles for specific p63 isoforms not only in maintaining epithelial stem cell populations, but also in cellular differentiation and neoplasia.     

IMP3 expression in biopsy specimens of colorectal cancer predicts lymph node metastasis and TNM stage

IMP3 is associated with lymph node metastasis and TNM stage and is a good independent prognostic biomarker for colorectal cancer (CRC). However, the expression status and clinical implication of IMP3 in biopsy specimens have not yet been studied. We aim to address whether the presence of IMP3 expression in preoperative biopsies of CRC could predict lymph node metastasis and TNM stage. In this study, we examined IMP3 expression in paired biopsy and resection specimens of 71 CRC and analyzed the correlation of IMP3 expression with clinicopathological parameters. In the biopsy specimens, IMP3 positive expression was observed in 56 of 71 cases (78.9%) whereas negative expression was observed in 15 of 71 cases (21.1%). In the resection specimens, IMP3 positive expression was detected in 83.1% cases (59/71) whereas negative expression was detected in 16.9% cases (12/71). The absolute concordance rate between biopsy and resection specimens was 90.1% (64/71). The Spearman correlation test documented the existence of a strong linear correlation between the percentage of IMP3-positive cells in the biopsy and resection specimen (r = 0.629; P < 0.001). IMP3 expression in resection specimens was significantly related to histological grade (P = 0.043), T classification (P = 0.035), lymph node metastasis (P = 0.023), TNM stage (P = 0.007), tumor border (P = 0.049) and tumor budding (P = 0.012). IMP3 expression in biopsy specimens was significantly related to lymph node metastasis (P = 0.004), TNM stage (P = 0.005) and tumor budding (P = 0.001). In conclusion, IMP3 expression in biopsy specimens could be used to predict lymph node metastasis and TNM stage in CRC patients.