Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

Immunotherapy for esophageal cancer:Where are we now and where can we go

Immune checkpoint inhibitor therapy has dramatically improved patient prognosis,and thereby transformed the treatment in various cancer types including esophageal squamous cell carcinoma(ESCC)in the past decade.Monoclonal antibodies that selectively inhibit programmed cell death-1(PD-1)activity has now become standard of care in the treatment of ESCC in metastatic settings,and has a high expectation to provide clinical benefit during perioperative period.Further,anti-cytotoxic T-lymphocyte–associated protein 4(CTLA-4)monoclonal antibody has also been approved in the treatment of recurrent/metastatic ESCC in combination with anti-PD-1 antibody.Well understanding of the existing evidence of immune-based treatments for ESCC,as well as recent clinical trials on various combinations with chemotherapy for different clinical settings including neoadjuvant,adjuvant,and metastatic diseases,may provide future prospects of ESCC treatment for better patient outcomes.

Effects of transferred NK4 gene on proliferation, migration, invasion and apoptosis of human prostate cancer DU145 cells

We investigated the ability of NK4, an antagonist of human hepatocyte growth factor （HGF）, to inhibit the influence of HGF on proliferation, migration, invasion and apoptosis of human prostate cancer cells. Expression vector pBudCE4.1-EGFP-NK4 containing NK4 cDNA was used to transfect human prostate cancer DU145 cells, and the effects of the autocrine NK4 on tumor cell proliferation, migration, invasion and apoptosis were assessed in vitro. in vivo, we subcutaneously implanted DU145 cells, mock-transfected clone （DU145/empty vector） cells and NK4- transfected clone （DU145/NK4） cells into nude mice, and then evaluated tumor growth, cell proliferation and cell apoptosis in vivo. We found that DU145/NK4 cells expressed NK4 protein. In the in vitro study, autocrine NK4 at- tenuated the HGF-induced tumor cell proliferation, migration and invasion, and stimulated apoptosis. Furthermore, autocrine NK4 effectively inhibited the HGF-induced phosphorylation of c-Met, extracellular signal-regulated kinase-1 （ERK1）. and protein kinase B 1/2 （Aktl/2）. Histological examination revealed that autocrine NK4 inhibited prolifera- tion and accelerated apoptosis of prostate cancer cells. These results show that genetic modification of DU145 cells with NK4 cDNA yields a significant effect on their proliferation, migration, invasion and apoptosis. Molecular targeting of HGF/c-Met by NK4 could be applied as a novel therapeutic approach to prostate cancer.

Diagnostic value of serum human epididymis protein 4 in esophageal squamous cell carcinoma

BACKGROUND Numerous studies have demonstrated that human epididymis protein 4(HE4)is overexpressed in various malignant tissues including ovarian,endometrial,lung,breast,pancreatic,and gastric cancers.However,no study has examined the diagnostic impact of HE4 in patient with esophageal squamous cell carcinoma(ESCC)until now.AIM To analyze the value of four serum tumor markers for the diagnosis of ESCC,and examine the associations of serum levels of HE4 with ESCC patients’clinicopathological characteristics.METHODS The case group consisted of 80 ESCC patients,which were compared to a control group of 56 patients with benign esophageal disease.Serum levels of HE4,carcinoma embryonic antigen(CEA),alpha fetal protein,and carbohydrate antigen 19-9(CA19-9)were detected by ELISA.The associations of serum HE4 levels with ESCC patients’clinicopathological characteristics such as gender,tumor location,and pathological stage were also examined after operation.RESULTS The result of ELISA showed that serum HE4 level was significantly higher in the patients with ESCC than in the controls,and the staining intensity was inversely correlated with the pathological T and N stages.Serum HE4 levels had a sensitivity of 66.2%and specificity of 78.6%when the cutoff value was set at 3.9 ng/mL.Moreover,the combined HE4 and CA19-9 increased the sensitivity to 83.33%,and interestingly,the combination of HE4 with CEA led to the most powerful sensitivity of 87.5%.Furthermore,A positive correlation was observed between HE4 serum levels and pathological T and N stages(P=0.0002 and 0.0017,respectively),but there was no correlation between HE4 serum levels and ESCC patient gender(P=0.4395)or tumor location(P=0.6777).CONCLUSION The results of this study suggest that detection of serum HE4 levels may be useful in auxiliary diagnosis and evaluation of the progression of ESCC.

Incidence of human papilloma virus in esophageal squamous cell carcinoma in patients from the Lublin region

AIM:To assess the prevalence of human papilloma virus(HPV) in esophageal squamous cell carcinoma(ESCC) in the south-eastern region of Poland.METHODS:The study population consisted of 56 ESCC patients and 35 controls.The controls were patients referred to our department due to other nonesophageal and non-oncological disorders with no gross or microscopic esophageal pathology as confirmed by endoscopy and histopathology.In the ESCC patients,samples were taken from normal mucosa(56 mucosa samples) and from the tumor(56 tumor samples).Tissue samples from the controls were taken from normal mucosa of the middle esophagus(35 control samples).Quantitative determination of DNA was carried out using a spectrophotometric method.Genomic DNA was isolated using the QIAamp DNA Midi Kit.HPV infection was identified following PCR amplification of the HPV gene sequence,using primers MY09 and MY11 complementary to the genome sequence of at least 33 types of HPV.The sequencing results were computationally analyzed using the basic local alignment search tool database.RESULTS:In tumor samples,HPV DNA was identified in 28 of 56 patients(50%).High risk HPV phenotypes(16 or/and 18) were found in 5 of 56 patients(8.9%),low risk in 19 of 56 patients(33.9%) and other types of HPV(37,81,97,CP6108) in 4 of 56 patients(7.1%).In mucosa samples,HPV DNA was isolated in 21 of 56 patients(37.5%).High risk HPV DNA was confirmed in 3 of 56 patients(5.3%),low risk HPV DNA in 12 of 56 patients(21.4%),and other types of HPV in 6 of 56 patients(10.7%).In control samples,HPV DNA was identified in 4 of 35 patients(11.4%) with no high risk HPV.The occurrence of HPV in ESCC patients was significantly higher than in the controls [28 of 56(50%) vs 4 of 35(11.4%),P < 0.001].In esophageal cancer patients,both in tumor and mucosa samples,the predominant HPV phenotypes were low risk HPV,isolated 4 times more frequently than high risk phenotypes [19 of 56(33.9%) vs 5 of 56(8.9%),P < 0.001].A higher prevalence of HPV was identified in female patients(71.4% vs 46.9%).Accordingly,the high risk phenotypes were isolated more frequently in female patients and this difference reached statistical significance [3 of 7(42.9%) vs 2 of 49(4.1%),P < 0.05].Of the pathological characteristics,only an infiltrative pattern of macroscopic tumor type significantly correlated with the presence of HPV DNA in ESCC samples [20 of 27(74.1%) vs 8 of 29(27.6%) for ulcerative or protruding macroscopic type,P < 0.05].The occurrence of total HPV DNA and both HPV high or low risk phenotypes did not significantly differ with regard to particular grades of cellular differentiation,phases in depth of tumor infiltration,grades of nodal involvement and stages of tumor progression.CONCLUSION:Low risk HPV phenotypes could be one of the co-activators or/and co-carcinogens in complex,progressive,multifactorial and multistep esophageal carcinogenesis.

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

AIM： To examine the presence of human papillomavirus （HPV） in esophageal squamous cell carcinoma （ESCC） specimens collected from Colombia and Chile located in the northern and southern ends of the continent, respectively.METHODS： We examined 47 and 26 formalin-fixed and paraffin-embedded ESCC specimens from Colombia and Chile, respectively. HPV was detected using GP5＋/GP6＋ primer pair for PCR, and confirmed by Southern blot analysis. Sequencing analysis of L1 region fragment was used to identify HPV genotype. In addition, P16^INK4A protein immunostaining of all the specimens was conducted.RESULTS： HPV was detected in 21 ESCC specimens （29%）. Sequencing analysis of L1 region fragment identified HPV-16 genome in 6 Colombian cases （13%） and in 5 Chilean cases （19%）. HPV-18 was detected in i0 cases （21%） in Colombia but not in any Chilean case. Since Chilean ESCC cases had a higher prevalence of HPV-16 （without statistical significance）, but a significantly lower prevalence of HPV-18 than in Colombian cases （P = 0.011） even though the two countries have similar ESCC incidence rates, the frequency of HPV-related ESCC may not be strongly affected by risk factors affecting the incidence of ESCC. HPV-16 genome was more frequently detected in p16 positive carcinomas, although the difference was not statistically significant. HPV-18 detection rate did not show any association with p16 expression. Well-differentiated tumors tended to have either HPV-16 or HPV-18 but the association was not statistically significant. HPV genotypes other than HPV-16 or 18 were not detected in either country.CONCLUSION： HPV-16 and HPV-18 genotypes can be found in ESCC specimens collected from two South American countries. Further studies on the relationship between HPV-16 presence and p16 expression in ESCC would aid understanding of the mechanism underlying the presence of HPV in ESCC.

Relationships of early esophageal cancer with human papillomavirus and alcohol metabolism

BACKGROUND It is well known that an alcohol consumption habit together with inactive heterozygous aldehyde dehydrogenase-2(ALDH2)is an important risk factor for the development of esophageal squamous cell carcinoma(ESCC).It remains controversial whether human papillomavirus(HPV)infection contributes to the occurrence/development of ESCC.There has been no study in which the relationship between ESCC and HPV in addition to alcohol dehydrogenase-1B(ADH1B)and ALDH2 genotypes was evaluated.AIM To evaluate relationships between HPV infection and development of esophageal cancer,particularly early esophageal cancer,based on ADH1B/ALDH2 polymorphisms.METHODS We conducted an exploratory retrospective study using new specimens,and we enrolled 145 patients who underwent endoscopic resection for superficial ESCC and had been observed for more than two years by both physical examination and endoscopic examination in Hokkaido University Hospital.Saliva was collected to analyze genetic polymorphisms of ADH1B/ALDH2.We performed in situ hybridization for resected specimens to detect HPV by using an HPV type 16/18 probe.RESULTS HPV was detected in 15(10.3%)of the 145 patients with ESCC.HPV-positive rates in inactive ALDH2*1/*2 and ALDH2*1/*1+*2/*2 were 10.8%and 9.8%,respectively(P=1.00).HPV-positive rates in slow-metabolizing ADH1B*1/*1 and ADH1B*1/*2+*2/*2 were 12.0%and 10.0%,respectively(P=0.72).HPV-positive rates in the heavy or moderate alcohol consumption group and the light or rare consumption group were 11.1%and 8.7%,respectively(P=0.68).HPV-positive rates in the heavy smoking group and the light or no smoking group were 11.8%and 8.3%,respectively(P=0.59).The 3-year incidence rates of secondary ESCC or head and neck cancer after initial treatment in the HPV-positive and HPVnegative groups were 14.4%and 21.4%(P=0.22),respectively.CONCLUSION In the present situation,HPV status is considered to be less important than other risk factors,such as alcohol consumption,smoking habit,ADH1B/ALDH2 polymorphisms,and HPV status would therefore have no effect on ESCC risk management.

No evidence of HPV DNA in esophageal squamous cell carcinoma in a population of Southern Brazil

AIM:To investigate the association between human papillomavirus(HPV)and esophageal squamous cell carcinoma(ESCC)in southern Brazil.METHODS:We studied 189 esophageal samples from125 patients from three different groups:(1)102 biopsies from 51 patients with ESCC,with one sample from the tumor and another from normal esophageal mucosa distant from the tumor;(2)50 esophageal biopsies from 37 patients with a previous diagnosis of head and neck squamous cell carcinoma(HNSCC);and(3)37 biopsies from esophageal mucosa with normal appearance from 37 dyspeptic patients,not exposed to smoking or alcohol consumption.Nested-polymerase chain reaction(PCR)with the MY09/11 and GP5/6 L1primers was used to detect HPV L1 in samples fixed in formalin and stored in paraffin blocks.All PCR reactions were performed with a positive control(cervicovaginal samples),with a negative control(Human Genomic DNA)and with a blank reaction containing all reagents except DNA.We took extreme care to prevent DNA contamination in sample collection,processing,and testing.RESULTS:The histological biopsies confirmed the diagnosis of ESCC in 52 samples(51 from ESCC group and 1 from the HNSCC group)and classified as well differentiated(12/52,23.1%),moderately differentiated(27/52,51.9%)or poorly differentiated(7/52,13.5%).One hundred twenty-eight esophageal biopsies were considered normal(51 from the ESCC group,42 from the HNSCC group and 35 from dyspeptic patients).Nine had esophagitis(7 from the HNSCC and 2 from dyspeptic patients).Of a total of 189 samples,only 6 samples had insufficient material for PCR analysis:1 from mucosa distant from the tumor in a patient with ESCC,3from patients with HNSCC and 2 from patients without cancer.In 183 samples(96.8%)GAPDH,G3PDH and/orβ-globin were amplified,thus indicating the adequacy of the DNA in those samples.HPV DNA was negative in all the 183 samples tested:52 with ESCC,9 with esophagitis and 122 with normal esophageal mucosa.CONCLUSION:There was no evidence of HPV infection in different ESCC from southern Brazil.

血清SCC-Ag、HE4、SMAD4水平与宫颈癌放化疗敏感性的相关性分析

目的分析血清鳞状细胞癌相关抗原(SCC-Ag)、人附睾分泌蛋白4(HE4)、SMAD4水平与宫颈癌放化疗敏感性的相关性。方法选取接受放化疗治疗的宫颈癌患者74例为研究对象,根据放化疗敏感性分为抵抗组和敏感组,分别有48例和26例。检测患者治疗前后血清SCC-Ag、HE4及SMAD4 mRNA水平,分析治疗前血清SCC-Ag、HE4、SMAD4水平与超声参数的相关性。绘制受试者工作特征曲线(ROC)描述治疗前SCC-Ag、HE4及SMAD4 mRNA水平对放化疗敏感性的预测价值。结果治疗后2组患者血清SCC-Ag、HE4水平较治疗前降低,SMAD4 mRNA水平较治疗前升高(P<0.05);且敏感组患者治疗前、后SCC-Ag、HE4水平均低于抵抗组,SMAD4 mRNA水平均高于抵抗组(P<0.05)。治疗前血清SCC-Ag、HE4与VI呈正相关(γ=0.518、0.544,P<0.05),SMAD4 mRNA与VI呈负相关(γ=-0.581,P<0.05)。ROC曲线显示,治疗前SCC-Ag、HE4及SMAD4 mRNA联合对放化疗敏感性的预测价值较高,AUC为0.863(95%CI:0.764~0.962)。结论血清SCC-Ag、HE4及SMAD4 mRNA水平与宫颈癌放化疗敏感性密切相关,且三者联合对放化疗敏感性具有较高的预测价值。

HPV18 E6 and E7 Intratumour Heterogeneity in Esophageal Cancer

The development of esophageal cancer accompanied by the presence of human papillomavirus (HPV) DNA into the host genome. By evaluating the expression of this virus for tumor cell origin and also their cell grows and migrations, we examined esophageal cancer clonality in the context of intra-tumor heterogeneity. In this research, we have checked the expression of HPV18 E6 and E7 in different single cell clones by the manual cell picking method in the HPV positive esophageal cancer (EC109), EC109 cell line used as a negative control, and Hela cell line used as the positive control. Quantitative real-time PCR (QRT-PCR) was run to detect the expression levels of HPV E6 and E7, Cell Counting Kit-8 (CCK-8) assay was used to examine cell proliferation, invasion assays performed using Costar chambers and wounding assay to study cell migrations in vitro. We investigated the intra-tumor heterogeneity of HPV E6 and E7 in esophageal cancer and the evaluation of the growth and migrations at the clonal level, using 10 single cell clones. In particular clones, C7 & C10 displayed a highly variable expression in both HPV E6 and E7 and weak in four clones (C1, C3, C4, and C9) consequently, the cell invasion, proliferation, and migration increase with increasing the level of HPV expression and inverse. In conclusion, the resulting based on single cell cloning showed the relationship between HPV and cell growth and migration in esophageal cancer. Future study in HPV DNA integration needed to explore the mains specific integration site of HPV DNA in esophageal cancer and molecular monitoring of the HPV for future prevention researches and also effective therapeutic strategies.

Effects of 4-(3-Chloro-Benzyl)-6,7-Dimethoxy-Quinazoline on Kinetics of P120-Catenin and Periplakin in Human Buccal Mucosa Squamous Carcinoma Cell Line

In order to detect molecular markers for the epidermal growth factor inhibitor 4-(3-chloro-benzyl)- 6,7-dimethoxy-quinazoline (tyrphostin), we investigated the kinetics of p120-catenin and periplakin in the human buccal mucosa squamous cancer cell line BICR 10 treated with 3 nM tyrphostin. Growth of BICR 10 cells was inhibited by treatment with tyrphostin. Although changes were not observed in the expression of EGFR and p120-catenin, expression of Akt, Src and periplakin in BICR 10 treated with 3 nM tyrphostin tended to decrease. In addition, phosphorylation of EGFR, Akt and Src was inhibited by treatment with tyrphostin. On immunocytochemical staining, immunoreactions with phosphorylated EGFR, phosphorylated Akt and phosphorylated p120-catenin were weak in BICR 10 treated with tyrphostin. There was a slight immunocy to chemical reaction to periplakin in BICR 10 cells induced by tyrphostin. In conclusion, the decrease in phosphorylation in EGFR and p120-catenin by tyrphostin, following the decrease in Src or Akt phosphorylation, may inhibit expression of several growth factors associated with the proliferation and migration of cancer cells.

血清HE4、SCC-Ag联合TCT检查诊断宫颈癌价值

目的:探讨血清人附睾上皮分泌蛋白4(HE4)、鳞状细胞癌相关抗原(SCC-Ag)联合宫颈薄层液基细胞学检测(TCT)诊断宫颈癌(CC)价值。方法:收集2021年10月-2023年7月本院病理确诊CC患者40例(CC组)、宫颈良性病变(CBL组,包括宫颈炎、宫颈上皮内瘤变)患者228例为CBL组。对比两组血清HE4、SCC-Ag水平,两组HE4、SCC-Ag、TCT检测CL阳性检出率;根据病理结果分析血清HE4、SCC-Ag水平、TCT诊断CL价值。结果:CC组血清HE4(188.71±39.15 pmol/L)、SCC-Ag(1.81±0.21 ng/ml)水平高于CBL组(73.65±11.52 pmol/L、0.64±0.12 ng/ml)(P<0.05)。TCT、HE4、SCC-Ag诊断CC的灵敏度(82.5%、72.5%、75.0%),特异度(77.6%、87.7%、86.8%),准确度(78.4%、85.5%、85.1%),3项指标联合诊断CC的灵敏度70.0%、特异度93.9%、准确度90.3%,联合检测准确度高于各单独指标检测(P<0.05)。结论:CC患者血清HE4、SCC-Ag水平及TCT检测阳性率均高于CBL患者,3项指标联合诊断准确度提高。

食管癌来源外泌体通过E2F4对CD4+T细胞向Th17细胞分化的影响

目的:研究食管癌来源外泌体对CD4+T细胞向Th17细胞分化的可能作用机制。方法:分离人正常食管上皮细胞HEEC和人食管癌细胞Eca-109来源外泌体(即HEEC-Exo、Eca-109-Exo),同时构建稳定表达的sh-NC和sh-E2F4的Eca-109细胞株并分离其外泌体(即sh-NC-Exo、sh-E2F4-Exo)。应用所得的外泌体处理CD4+T细胞;此外,应用稳定表达sh-NC和sh-E2F4的Eca-109细胞株建立食管癌异种移植小鼠模型。流式细胞术检测CD4+T细胞中Th17细胞比例;qRT-PCR检测E2F转录因子4(E2F4)mRNA表达;Western blot检测E2F4、维甲酸相关核孤儿受体γt(ROR-γt)和IL-17蛋白表达;ELISA试剂盒检测IL-17的含量。结果:与PBS组相比,HEEC-Exo组CD4+T细胞中Th17细胞比例、ROR-γt蛋白的表达、IL-17含量以及E2F4mRNA和蛋白表达差异无统计学意义(P>0.05);与HEEC-Exo组相比,Eca-109-Exo组CD4+T细胞中Th17细胞比例、ROR-γt蛋白表达、IL-17含量以及E2F4 mRNA和蛋白表达明显升高(P<0.05)。与Eca-109-Exo组相比,sh-NC-Exo组CD4+T细胞中Th17细胞比例、ROR-γt蛋白表达、IL-17含量以及E2F4 mRNA和蛋白表达差异无统计学意义(P>0.05);与sh-NC-Exo组相比,shE2F4-Exo组CD4+T细胞中Th17细胞比例、ROR-γt蛋白表达、IL-17含量以及E2F4 mRNA和蛋白表达均明显降低(P<0.05)。与Eca-109组相比,sh-NC组小鼠肿瘤大小和E2F4、ROR-γt、IL-17蛋白表达差异无统计学意义(P>0.05);与sh-NC组相比,sh-E2F4组小鼠肿瘤大小和E2F4、ROR-γt、IL-17蛋白表达均明显降低(P<0.05)。结论:食管癌来源外泌体可能通过E2F4促进CD4+T细胞向Th17细胞分化,从而促进肿瘤生长。

血清HE4、CEA及ESM-1联合检测对肺癌的诊断价值

目的探讨人血清中附睾蛋白4(humanepididymis 4,HE4)、癌胚抗原(carcinoembryonic antigen,CEA)及内皮细胞特异性分子-1(endothelial cell specific molecule-1,ESM-1)三项检验指标在肺癌诊断中的价值。方法采用化学发光法检测50例肺癌患者(肺癌组)、50例肺部良性病变患者(良性组)血清中HE4、CEA水平,应用ELISA法检测ESM-1水平,分析比较2组三项指标水平差异,采用ROC曲线比较HE4、CEA及ESM-1单项与联合检测对肺癌的诊断性能。结果HE4、CEA、ESM-1的检测水平肺癌组明显高于良性组,差异均有统计学意义(Z=-2.292,P=0.022;Z=-5.412,P=0.000;Z=-6.431,P=0.000);ESM-1的ROC曲线下面积(0.983)>CEA(0.907)>HE4(0.672),结果表明在诊断肺癌中,ESM-1ROC面积最大,有较高的诊断意义;ESM-1水平在26.31 ng/ml时,诊断肺癌的敏感度为96.7%,特异度为96.7%;通过绘制ROC曲线发现AUC CEA+HE4+ESM-1=AUC CEA+ESM-1>AUC HE4+ESM-1>AUC CEA+HE4,通过Z检验分析,发现CEA+ESM-1联合检测与CEA+HE4+ESM-1检测的ROC曲线下面积均为0.997,无统计学意义(P>0.05),两者诊断效率相当。结论CEA和ESM-1对肺癌的诊断有较高应用价值,CEA+ESM-1联合检测可使肺癌在诊断效率上得到显著提高。

血清CYFRA21-1、CEA、SCC、HE4、ProGRP对肺癌早期诊断及病理类型鉴别的临床价值

目的探究肿瘤标志物血清细胞角蛋白19片段(CYFRA21-1)、癌胚抗原(CEA)、鳞状细胞癌抗原(SCC)、人附睾蛋白4(HE4)、胃泌素释放肽前体(ProGRP)对肺癌早期诊断及病理类型鉴别的临床价值。方法选取2021年2月至2022年2月安康市中心医院收治的280例肺癌早期患者作为肺癌组,同期收治的100例肺部良性疾病患者作为良性对照组,比较两组患者的血清CYFRA21-1、CEA、SCC、HE4及ProGRP水平,采用受试者工作特征曲线(ROC)分析血清CYFRA21-1、CEA、SCC、HE4、Pro GRP以及联合检测对肺癌早期诊断的临床价值,并根据肺癌组患者的病理分型结果比较非小细胞肺癌(NSCLC)与小细胞肺癌(SCLC)患者的血清CYFRA21-1、CEA、SCC、HE4及ProGRP水平。结果肺癌组患者的血清CYFRA21-1、CEA、SCC、HE4及ProGRP水平分别为(2.04±1.87)ng/mL、(3.08±0.82)ng/mL、(4.51±1.54)ng/mL、(60.14±15.88)pmol/L、(61.27±19.34)μg/L,明显高于良性对照组的(1.15±0.43)ng/mL、(2.54±0.71)ng/mL、(3.09±1.68)ng/mL、(45.14±17.56)pmol/L、(50.14±18.73)μg/L,差异均有统计学意义(P<0.05);经ROC分析结果显示,血清CYFRA21-1诊断肺癌早期的最佳截断值为1.830 ng/m L,CEA为2.856 ng/mL,SCC为3.140 ng/mL,HE4为50.340 pmol/L,ProGRP为57.605μg/L,5项指标联合诊断肺癌早期的AUC及敏感度达到0.901、89.30%,均高于单一指标诊断,差异有统计学意义(P<0.05);SCLC组患者的ProGRP水平明显高于NSCLC组,差异有统计学意义(P<0.05),而两组患者的CYFRA21-1、CEA、SCC及HE4水平比较差异均无统计学意义(P>0.05);腺癌组患者的SCC为(5.50±2.73)ng/mL,明显高于鳞癌组的(1.42±0.05)ng/mL,差异有统计学意义(P<0.05),而两组患者的CYFRA21-1、CEA、HE4及ProGRP水平比较差异均无统计学意义(P>0.05)。结论与肺部良性疾病患者比较,肺癌早期患者血清CYFRA21-1、CEA、SCC、HE4及ProGRP水平更高,且其联合检测诊断早期肺癌的临床价值高,血清ProGRP鉴别NSCLC与SCLC,血清SCC鉴别腺癌与鳞癌均有一定价值,建议临床密切监测。

宫颈癌患者OCT4与临床病理、HPV感染及SCC-Ag水平的关系

目的 研究宫颈癌中八聚体结合转录因子4(OCT4)的表达与临床病理、人乳头状瘤病毒(HPV)感染及鳞状细胞癌抗原(SCC-Ag)水平的关系。方法 选取2018年1月至2021年10月来永城市人民医院接受治疗的150例宫颈癌患者的包埋肿瘤组织标本,收集患者分期、分化、类型、淋巴结转移等信息,以免疫组织化学检测试剂盒检测OCT4,以免疫组织化学检测HPV,以化学发光免疫分析法测定试剂盒检测SCC-Ag。结果 OCT4与类型、分期、分化、淋巴结转移之间差异无统计学意义(P>0.05)。HPV感染情况不同的患者及SCC-Ag表达水平不同的患者,OCT4表达差异存在统计学意义(P<0.05)。结论 宫颈癌患者中,OCT4表达在不同临床病理中无差异,与HPV感染及SCC-Ag表达相关。

Value of serum NGAL and HE4 content change for diagnosing lung cancer and evaluating disease progression

Objective: To study the value of serum NGAL and HE4 content change for diagnosing non-small cell lung cancer (NSCLC) and evaluating disease progression. Methods: Patients who were diagnosed with NSCLC and underwent surgical resection in the First Affiliated Hospital of Chengdu Medical College between February 2015 and March 2017 were selected as the NSCLC group, and healthy volunteers who received physical examination over the same period were selected as control group. The NGAL and HE4 contents in serum of NSCLC group and control group as well as the expression of proliferation, apoptosis and invasion genes in NSCLC lesion of NSCLC group were detected. Results: Serum NGAL and HE4 levels of NSCLC group were significantly higher than those of control group, serum NGAL and HE4 levels of patients with TNM II and TNM III NSCLC were significantly higher than those of patients with TNM I NSCLC, and serum NGAL and HE4 levels of patients with TNM III NSCLC were significantly higher than those of patients with TNM II NSCLC;c-myc, Survivin, CatL, Vimentin, Slug and Snail mRNA expression in NSCLC lesion were significantly higher than those in adjacent lesion and positively correlated with serum NGAL and HE4 levels while PTEN, LATS1 and E-cadherin mRNA expression were significantly lower than those in adjacent lesion and negatively correlated with serum NGAL and HE4 levels. Conclusion: Abnormally elevated NGAL and HE4 in the serum of NSCLC patients can evaluate the proliferation and invasion of cancer cells to a certain extent.

BRD4通过靶向GSTP1调控食管癌TE-1细胞顺铂敏感性

目的揭示食管癌组织中溴结构域蛋白4(BRD4)表达水平,并分析其与谷胱甘肽硫基转移酶P1(GSTP1)分子的联系,探究BRD4在人食管癌细胞(TE-1细胞)顺铂敏感性调控中的作用。方法基于癌症基因组图谱(TCGA)数据利用R语言包解析BRD4在食管癌及癌旁正常组织中表达情况;基因表法普交互分析(GEPIA2)在线评估食管癌肿瘤组织中BRD4与化疗药物解毒酶GSTP1表达的联系;通过小干扰RNA(siRNA)和靶向抑制剂(+)-JQ1(25、50和100 nM)分别处理食管癌TE-1细胞,通过蛋白免疫印迹法(Western blot)验证食管癌TE-1细胞中BRD4及GSTP1蛋白表达变化;TE-1细胞(+)-JQ1预处理(50 nM,24 h)前后及结合顺铂(DDP,5μg/ml,24 h),运用Western blot实验检测凋亡相关蛋白Bcl-2关联X蛋白(Bax)、淋巴细胞瘤-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)及半胱氨酸天冬氨酸蛋白酶3切割体(cleaved-caspase-3)表达变化;采用CCK-8法检测TE-1细胞经(+)-JQ1(50 nM,24 h)抑制BRD4作用后DDP敏感性的改变。结果TCGA数据解析后结果显示,食管癌肿瘤组织中BRD4的mRNA总体表达水平相较于癌旁正常组织有明显升高[(5.61±0.57)VS(4.80±0.59)],差异有统计意义(P<0.001)。进一步通过对比配对肿瘤和癌旁正常组织样本发现,食管癌患者食管癌组织中BRD4表达水平(5.57±0.31)普遍显著高于其癌旁正常组织的(4.77±0.44),差异有统计学意义(P<0.01)。GEPIA2数据库发现,在食管癌化疗药物耐药形成过程中关键解毒结合酶GSTP1与肿瘤组织中BRD4表达呈正相关(r=0.26,P=0.00029<0.001)。结合靶向siRNA处理Western blot结果显示,siRNA-BRD4处理后食管癌TE-1细胞中BRD4和GSTP1的表达水平较NC组均有明显升高,而siRNA-GSTP1后TE-1细胞中GSTP1表达下调未对BRD4蛋白产生明显影响。进一步采取BRD4靶向抑制剂的梯度浓度处理证实,伴随(+)-JQ1浓度升高TE-1细胞中GSTP1蛋白的表达水平较阴性对照组出现明显下降趋势,而由于(+)-JQ1主要是通过结合于BRD4的溴结构域发挥作用,仅在最高浓度处理(100 nM)组食管癌细胞中BRD4出现表达抑制作用。低浓度的(+)-JQ1(50 nM)处理可引起食管癌TE-1细胞抗凋亡蛋白Bcl-2的表达下调以及凋亡相关蛋白Bax表达上升,但对caspase-3并无明显影响,而在与DDP联合应用时可见caspase-3的活化形式cleaved-caspase-3的相比DDP单独使用组细胞有显著表达上调。Western blot结果表明,(+)-JQ1预处理抑制BRD4的功能可以提高DDP对食管癌细胞的杀伤作用。CCK-8结果显示,食管癌TE-1细胞经抑制BRD4蛋白功能[(+)-JQ1,50 nM,24 h]预处理后可显著提高对化疗药物DDP的敏感性,DMSO对照组DDP的半数抑制浓度(IC50)值为(19.43±0.59)μg/ml,而(+)-JQ1预处理后降至(10.46±0.24)μg/ml,差异有统计学意义(P<0.05)。结论食管癌中高表达的BRD4可能通过GSTP1分子参与DDP敏感性调控。

宫颈癌患者血清HE4、CEACAM6、AFP表达水平及其与临床病理特征的相关性

目的探究宫颈癌患者血清人附睾蛋白4(HE4)、癌胚抗原相关细胞黏附分子6(CEACAM6)、甲胎蛋白(AFP)表达水平及其与临床病理特征的相关性。方法选取2020年5月至2022年10月南阳南石医院妇产科收治的92例宫颈癌患者作为宫颈癌组,并选择同期46例宫颈上皮内瘤变患者作为良性组,46例健康体检者作为对照组。比较三组受检者的血清HE4、CEACAM6、AFP水平;采用Spearson相关性检验分析血清HE4、CEACAM6、AFP水平与临床病理特征相关性。结果宫颈癌组患者的血清HE4、CEACAM6、AFP水平分别为(124.79±15.35)pmol/L、(78.15±11.24)mg/L、(24.11±3.65)ng/mL,明显高于对照组的(37.54±3.11)pmol/L、(15.64±3.21)mg/L、(6.35±1.49)ng/mL和良性组的(75.61±8.57)pmol/L、(30.23±4.55)mg/L、(30.23±4.55)ng/mL,差异均有统计学意义(均P<0.001);不同淋巴结转移、分化程度、临床分期、浸润深度宫颈癌患者血清HE4、CEACAM6、AFP表达水平比较差异均有统计学意义(P<0.001);经Spearson相关性分析结果显示,血清HE4、CEACAM6、AFP与淋巴结转移、临床分期、浸润深度呈正相关(均P<0.001),与分化程度呈负相关(均P<0.001)。结论宫颈癌患者血清内HE4、CEACAM6、AFP呈高水平状态,且与宫颈癌患者的临床病理特征密切相关,可为宫颈癌的早期诊断提供参考依据。

血清人附睾蛋白4对非小细胞肺癌的诊断价值

目的探讨人附睾蛋白4(HE4)在非小细胞肺癌(NSCLC)患者血清中的表达及在NSCLC诊断中的价值。方法选取97例NSCLC患者作为肺癌组,47例健康体检人群作为对照组。测定所有研究对象血清HE4、癌胚抗原(CEA)、糖类抗原125(CA125)、糖类抗原19-9(CA19-9)水平,评价其对肺癌的诊断效能。结果肺癌组患者血清HE4、CEA、CA19-9、CA125水平均明显高于对照组,差异均有统计学意义(P﹤0.01)。男性NSCLC患者血清HE4水平高于女性患者,差异有统计学意义(P﹤0.05)。有远处转移的NSCLC患者血清HE4、CEA、CA19-9、CA125水平均明显高于无远处转移患者,差异均有统计学意义(P﹤0.01)。受试者工作特征(ROC)曲线分析显示,HE4联合CEA或CA125检测诊断肺癌的曲线下面积(AUC)均大于HE4单独检测,联合检测对肺癌诊断有较高的特异度。结论HE4对肺癌有一定的辅助诊断价值,与CA125或CEA联合检测对NSCLC诊断的特异度高于HE4单独检测,可提高肺癌的诊断率,具有较高的临床应用价值。