粪便SDC2基因甲基化检测联合FOB、 CEA和CA199检测在结直肠癌早期筛查中的应用

目的:探讨粪便黏结蛋白聚糖2(SDC2)基因甲基化检测联合粪便潜血(FOB)、癌胚抗原(CEA)和糖类抗原199(CA199)检测在结直肠癌早期筛查中的应用价值。方法:从本院采集的358例受试者粪便样本中,实施粪便SDC2基因甲基化检测,并对结果进行深入分析。进一步筛选出符合入组条件的160例患者,分为健康对照组、疾病对照组、癌前病变组和结直肠癌组,每组各40例。补充完善血清CEA、CA199及FOB检测,统计并评估各项指标的诊断效能。结果:与健康对照组及疾病对照组对比,结直肠癌组及癌前病变组的粪便SDC2基因甲基化水平、FOB阳性率以及CEA、CA199的表达量均呈现显著性上升(P<0.05)。其中,结直肠癌组粪便SDC2基因甲基化检测的阳性率最高,为85.0%;其次为粪便潜血试验(72.5%)。这两项指标明显高于CEA和CA199的检测阳性率。针对不同特征人群进行粪便SDC2基因筛查时发现,阳性率与受检者的年龄、症状表现及其地域来源存在关联性,但与性别并无显著相关性。结论:粪便SDC2基因甲基化检测是一种简便快捷、无创且灵敏度较高的新型检测方式,能很大程度上提高早期结直肠癌及...     

大肠癌血液诊断标志物的研究进展

结直肠癌(CRC)血液标志物主要分为4类：核酸(RNA/DNA/信使RNA/microRNA)、细胞因子、抗体和蛋白质。在核酸中最有希望的血液标志物是NEAT_v2非编码RNA,SDC2甲基化和Septin 9甲基化。检测结直肠癌的最有希望的细胞因子是白细胞介素(IL)-8,最有希望的蛋白类标志物是CA11-19糖蛋白...     

粪便TFPI-2甲基化联合外周血内糖类抗原199在老年直肠癌筛查中的应用价值

目的:分析粪便组织因子途径抑制物-2(TFPI-2)甲基化联合外周血内糖类抗原199(CA199)在老年直肠癌筛查中的应用价值。方法:选择我院2020-2021年诊治的老年直肠癌患者72例,将其纳入观察组;另在同期保证性别和年龄资料均衡情况下选择疑似直肠癌但证实为健康受试者72名,将其纳入对照组。检查粪便TFPI-2甲基化、外周血内CA199阳性情况,分析联合诊断价值。结果:观察组患者粪便TFPI-2甲基化、CA199阳性率分别高于对照组(P<0.05)。粪便TFPI-2甲基化联合CA199检测直肠癌诊断效能分别为灵敏度94.57%、特异度77.26%、约登指数0.721、AUC0.863,灵敏度、约登指数及AUC最大。结论:粪便TFPI-2甲基化、外周血内CA199联合检测可以做到优势互补,提高直肠癌诊断效能,建议推广应用。     

粪便p16基因甲基化状态在结直肠癌早期诊断的意义

目的探索p16基因甲基化状态与结直肠癌（CRC）早期诊断的可行性。方法从61例CRC患者、21例腺瘤患者和20例正常对照者粪便中提取DNA,采用巢式甲基化特异性PCR（nMSP）技术分析其p16基因甲基化改变状态。结果CRC、腺瘤、正常对照者粪便标本p16基因甲基化检出率分别为77．0％（47／61）、59．3％（16／27）、5．0％（1／20）。检测p16基因甲基化诊断CRC的敏感性和特异性分别为71．6％和95．0％。p16基因甲基化的阳性状态与CRC患者年龄、性别、肿瘤发生部位无明显关系,p16基因甲基化的阳性状态与肿瘤的Dukes分期、组织分化程度和淋巴结转移有关。结论p16基因甲基化改变发生在CRC形成的早期阶段,检测粪便DNA中p16基因的甲基化状态有望成为CRC早期无创诊断及CRC高风险人群筛查诊断的新途径。     

三甲基转移酶SETD2表达下调促进结肠直肠癌细胞的迁移和增殖

目的:研究组蛋白H3K36三甲基转移酶SETD2对结肠直肠癌(CRC)细胞发生和生长的影响。方法:通过短发夹RNA转染降低CRC细胞系HCT116和SW480中SETD2的mRNA和蛋白质表达水平。检测转染后的CRC细胞系中三甲基化H3K36的蛋白质表达水平和肿瘤细胞增殖及迁移能力的变化,通过裸鼠成瘤实验观察KD组和对照组SETD2对于肿瘤生长的影响。结果:在CRC细胞中,随着SETD2的mRNA及蛋白质表达水平下降,检测到三甲基化H3K36蛋白质表达下降,且CRC细胞的增殖和迁移能力显著提升(P0.05)。在裸鼠成瘤实验中通过免疫组织化学染色法,检测到KD组的小鼠肿瘤组织内三甲基化H3K36的蛋白质表达水平明显低于对照组。KD组裸鼠肿瘤的发生时间明显早于对照组,肿瘤的直径也明显大于对照组(P0.01)。结论:下调SETD2的mRNA和蛋白质表达导致肿瘤组织三甲基化H3K36蛋白质表达下降,迁移和增殖能力增强,研究提示SETD2在CRC发生中的重要作用。     

基于骨组织Dlx5启动子甲基化探究补肾中药复方对去卵巢骨质疏松症大鼠的疗效机制

目的基于骨组织Dlx5启动子甲基化水平,探索补肾中药复方对大鼠去卵巢骨质疏松症的疗效机制。方法去卵巢建立绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠模型,分为正常组、模型组、补肾中药复方组、仙灵骨葆阳性对照组。灌胃14周后,X线吸收测量法检测骨密度,质谱法检测Dlx5启动子甲基化水平。结果与正常组比较,模型组第1~6腰椎骨密度明显降低(P<0.01),骨组织Dlx5启动子-470 bp^-4 bp CpG2.3甲基化水平明显升高(P<0.01)。②与模型组比较,补肾中药复方组、仙灵骨葆阳性对照组第1~6腰椎骨密度明显升高(P<0.05),骨组织Dlx5启动子-470 bp^-4 bp CpG1甲基化水平明显降低(P<0.05)。结论补肾中药复方通过降低骨组织Dlx5启动子甲基化水平的表观遗传学机制,有效防治PMOP。     

BNIP3基因甲基化与早期结直肠癌患者临床病理特征的关系

目的分析早期结直肠癌组织中BNIP3基因启动子区CpG岛甲基化与患者临床病理特征的关系,评价其在结直肠癌发生发展中的作用。方法用甲基化特异性PCR技术分析107例早期结直肠癌患者肿瘤组织中BNIP3基因启动子区CpG岛甲基化状态。结果本组结直肠癌组织标本中BNIP3基因启动子区CpG岛甲基化阳性率为58.9%(63/107)。右半结肠组肿瘤组织中BNIP3基因甲基化明显高于左半结肠组(P<0.001);低分化组明显高于高中分化组(P=0.002)。BNIP3基因甲基化在患者性别、年龄、肿瘤大小和分期等分组间无明显差异。结论早期结直肠癌组织中存在BNIP3基因启动子区CpG岛甲基化。BNIP3基因高甲基化与早期结直肠癌部位和分化程度密切相关。     

肝细胞癌组织中Ras相关区域家族蛋白1A、肿瘤高甲基化基因1和p73基因的异常甲基化

目的探讨肝细胞癌中RAS相关区域家族蛋白1A(RASSF1A)、肿瘤高甲基化基因1(H IC1)和P73基因启动子异常甲基化状况及其与患者临床和病理因素之间的关系。方法收集肝细胞癌组织标本和对应的远癌组织标本各40例及2例健康人的肝组织标本;通过甲基化特异的聚合酶链反应方法检测这些标本中RASSF1A、H IC1和P73基因启动子区的甲基化状况。结果RASSF1A基因在癌和远癌组织中启动子区的甲基化率分别为90.0%和72.5%,癌组织中的甲基化率高于远癌组织(P<0.05)。H IC1基因在癌和远癌组织中的甲基化率分别为77.5%和70.0%。P73基因在癌组织中甲基化率为5.0%,在远癌组织没有发现甲基化。健康人肝组织中未发现任何上述基因的异常甲基化。远癌组织中H IC1基因甲基化患者的年龄[(48±14)岁]比无甲基化患者[(57±11)岁]小(T=2.049,P=0.047)。3个基因甲基化发生情况与患者性别、是否合并乙型肝炎、肝硬化、甲胎蛋白水平、肿瘤大小、有无包膜和门静脉癌栓以及TNM分期之间未发现有统计学相关性。结论肝细胞癌中RASSF1A和H IC1基因启动子的高甲基化是普遍现象,年轻肝细胞癌患者H IC1甲基化现象更常见。     

结直肠癌患者分泌型卷曲相关蛋白2基因超甲基化研究

目的 分析结直肠癌患者粪便和外周血中分泌型卷曲相关蛋白2(SFRF2)基因甲基化与结直肠癌Dukes分期及组织学分型的关系,评价用SFRP2基因筛查结直肠癌和判断病情的价值.方法 用甲基化特异性PCR技术分析169例结直肠癌,63例结肠腺瘤和46例非腺瘤性息肉患者的瘤组织、粪便和外周血中SFRY2基因甲基化状态,以30例健康志愿者肠镜下肠黏膜活性组织作为对照. 结果本组SFRP2基因在88.2%的结直肠癌组织,65.1%的腺瘤组织和45.7%的非腺瘤性息肉组织中发生超甲基化;在相应的粪便中分别有84.0%、46.0%和32.6%发生超甲基化;在相应的外周血中分别有66.9%、6.4%和2.2%发生超甲基化.在正常对照的结肠黏膜组织和外周血中未检测到SFRP2基因超甲基化,但在粪便中有2例SFRIY2基因甲基化.此外,肿瘤组织和粪便中SFRP2与肿瘤的临床特征包括性别、年龄、位置、大小、Dukes分期等无显著相关性,与分化和浸润程度呈正相关,而外周血中SFRP2基因超甲基化与Dukes分期呈正相关.结论 粪便中SFRF2基因超甲基化可以作为结直肠癌筛查的一种新型分子标记物;外周血中SFBP2基因超甲基化与肿瘤分期呈正相关,可能对判定结直肠癌恶性程度和预后有一定的意义.     

结直肠癌患者分泌型卷曲相关蛋白2基因超甲基化研究

目的 分析结直肠癌患者粪便和外周血中分泌型卷曲相关蛋白2(SFRF2)基因甲基化与结直肠癌Dukes分期及组织学分型的关系,评价用SFRP2基因筛查结直肠癌和判断病情的价值.方法 用甲基化特异性PCR技术分析169例结直肠癌,63例结肠腺瘤和46例非腺瘤性息肉患者的瘤组织、粪便和外周血中SFRY2基因甲基化状态,以30例健康志愿者肠镜下肠黏膜活性组织作为对照. 结果本组SFRP2基因在88.2%的结直肠癌组织,65.1%的腺瘤组织和45.7%的非腺瘤性息肉组织中发生超甲基化;在相应的粪便中分别有84.0%、46.0%和32.6%发生超甲基化;在相应的外周血中分别有66.9%、6.4%和2.2%发生超甲基化.在正常对照的结肠黏膜组织和外周血中未检测到SFRP2基因超甲基化,但在粪便中有2例SFRIY2基因甲基化.此外,肿瘤组织和粪便中SFRP2与肿瘤的临床特征包括性别、年龄、位置、大小、Dukes分期等无显著相关性,与分化和浸润程度呈正相关,而外周血中SFRP2基因超甲基化与Dukes分期呈正相关.结论 粪便中SFRF2基因超甲基化可以作为结直肠癌筛查的一种新型分子标记物;外周血中SFBP2基因超甲基化与肿瘤分期呈正相关,可能对判定结直肠癌恶性程度和预后有一定的意义.     

Impact of faecal haemoglobin based triage of bowel symptoms presenting to primary care on colorectal cancer diagnosis

Aim

Faecal immunochemical testing (FIT) for faecal haemoglobin was introduced into primary care in National Health Service Tayside in 2015 as an adjunct to clinical assessment of new bowel symptoms. We aimed to assess the impact of FIT-based triage in primary care on colorectal cancer (CRC) diagnosis.

Method

Cancer audit data between January 2016 and December 2019 were reviewed to identify all patients diagnosed locally with CRC. The mode of presentation and stage at diagnosis were noted and patient records were interrogated to identify whether FIT and full blood count (FBC) had been performed prior to referral. Results were compared between the FIT and non-FIT groups.

Results

In all, 1245 patients were diagnosed with CRC of whom 581 (46.7%) presented through primary care. FIT was performed prior to referral in 440/581 (75.7%), with the proportion increasing from 62.3% in 2016 to 85.8% in 2019. At faecal haemoglobin ≥10 μg Hb/g faeces, sensitivity for CRC was 94.1%. Over the study period the annual proportion of non-emergency presentations increased significantly; presentations from primary care increased from 43.1% to 53.5% (P = 0.0096). After excluding non-FIT patients who had an overt CRC at referral, there was no difference in stage at diagnosis between FIT and non-FIT cancers. Safety-netting with FBC was widely used in our cohort with 97.3% of FIT patients having also had FBC.

Conclusion

FIT-based triage of new bowel symptoms in primary care is associated with increased non-emergency presentation of CRC but this did not influence stage at diagnosis.     

乳腺癌组织中AXIN2基因启动子区甲基化表达及临床意义

目的本研究旨在通过检测乳腺癌组织中AXIN2基因启动子区甲基化状态,分析其与乳腺癌相关危险因素、临床病理特征及mRNA表达间的关系,探讨其在临床应用中的价值。方法选取2018年1月至2019年12月在山东第一医科大学附属肥城市人民医院普外科经手术切除治疗的84例女性乳腺癌患者的乳腺癌组织和癌旁组织,检测组织的AXIN2基因启动子的甲基化状态和mRNA表达量,使用不同浓度的甲基转移酶抑制剂(5-Azacytidine,5-Aza)处理人乳腺癌ZR-75-1细胞,且进行乳腺癌相关危险因素和临床病理特征的收集与分析。结果乳腺癌组织的AXIN2基因启动子甲基化阳性率(40.5%)显著高于癌旁组织的甲基化阳性率(19.0%)(χ2=10.401,P=0.001)。AXIN2甲基化阳性率与乳腺癌患者的初产年龄(χ2=5.467,P=0.019)、是否有人工流产史(χ2=8.372,P=0.006)有关;AXIN2甲基化阳性率与是否发生淋巴结转移(χ2=5.338,P=0.021)、ER表达状态(χ2=9.141,P=0.002)及PR表达状态(χ2=6.918,P=0.009)有关。此外,乳腺癌组织的AXIN2基因mRNA相对表达量(00.22±0.03)显著低于癌旁组织的mRNA相对表达量(0.46±0.06)(t=3.527,P<0.001);甲基化阳性乳腺癌组织的mRNA相对表达量(0.13±0.02)显著低于甲基化阴性乳腺癌组织的mRNA相对表达量(0.29±0.04)(t=3.616,P<0.001)。ZR-75-1细胞使用5-Aza处理后,AXIN2基因mRNA相对表达量显著升高(t=3.824,P<0.001)。结论AXIN2基因启动子区DNA甲基化与乳腺癌发生机制有关,在临床中有一定应用价值。     

Clinical Significance of Large Tenascin-C Spliced Variant as a Potential Biomarker for Colorectal Cancer

Background Tenascin-C is an extracellular matrix protein forming various types of spliced variants. Low molecule variants are transiently present, but large spliced variants are predominantly overexpressed in proliferative processes or tumorigenesis in some varieties of cancer. However, the detection of the plasma level of large tenascin-C spliced variant (L-Tn-CSV) in colorectal cancer (CRC) has not been clarified. This study was performed to validate elevated plasma L-Tn-CSV levels as a possible biomarker for CRC. Materials and Methods Plasma samples were obtained before resection and from time to time postoperatively and stored at −80°C until assay. Plasma L-Tn-CSV levels were evaluated in patients with primary (n = 162) and with recurrent (n = 20) CRC, including 48 healthy volunteers, measured by ELISA. Results The average plasma L-Tn-CSV concentrations of patients with primary CRC were 5,260 ± 3,243.3 pg/ml and of patients with recurrent CRC 4,106 ± 2,261.1 pg/ml, which were significantly elevated in comparison with those of healthy volunteers (2,364.3 ± 7,49.6). The sensitivity for detecting CRC using plasma L-Tn-CSV was 56.6%, based on the mean ± 2 SD of the concentrations of healthy controls (3,863.5), which was significantly higher than CEA (40.1%) and CA19-9 (23.6%). No obvious associations were evident between plasma L-Tn-CSV status and values of CEA and CA19-9 respectively. Statistically significant differences in plasma L-Tn-CSV were observed depending on tumor depth, lymph node metastasis, and TNM stage. Negative conversions of plasma L-Tn-CSV levels 6 months after resection were significantly higher in the completely curative resection group than in the non-curative groups (P < 0.001). Conclusion The plasma L-Tn-CSV may serve very well as a useful biomarker for tumor staging and postoperative monitoring of preoperatively positive CRC that is independent and exceeds conventional tumor markers.     

Phenotype of microsatellite-stable colorectal carcinomas with CpG island methylation

A distinctive pathway of colorectal carcinogenesis termed CpG island methylator phenotype is characterized by extensive DNA methylation in colorectal carcinoma (CRC) cells but not in nonneoplastic mucosa. Many CRCs with CpG island methylator phenotype have methylation of the hMLH1 mismatch repair gene and consequently have high levels of microsatellite instability (MSI-H). MSI-H confers distinctive clinical-pathologic features, but the phenotype of microsatellite-stable CRC with methylation has not been characterized in detail. We therefore examined the clinical-pathologic features of 87 sporadic microsatellite-stable CRCs that had been characterized for methylation of p16, p14, MGMT, hMLH1, MINT1, MINT2, and MINT31. Regression analyses of each clinical-pathologic characteristic were run against the individual and aggregated methylation markers to evaluate and quantify associations. CpG island methylation was associated with right-sided carcinoma (odds ratio = 6.9, P = 0.03). Paucity of gland formation, indicating poor differentiation, was strongly associated with methylation (beta = -42.6, P = 0.0008), as were presence of cribriform glands (beta = 34.3, P = 0.02) and lack of corkscrew/serrated glandular pattern (beta = -32.5, P = 0.03). Our epigenotype-phenotype correlation study shows that microsatellite-stable CRC with CpG island methylation have a distinctive pathologic phenotype with both similarities to and differences from MSI-H tumors.     

GP access to FIT increases the proportion of colorectal cancers detected on urgent pathways in symptomatic patients in Nottingham

ObjectiveService evaluation of GP access to Faecal Immunochemical Test (FIT) for colorectal cancer (CRC) detection in Nottinghamshire and use of FIT for “rule out”, “rule in” and “first test selection”.DesignRetrospective audit of FIT results, CRC outcomes and resource utilisation before and after introduction of FIT in Primary Care in November 2017. Data from the new pathway up to December 2018 was compared with previous experience.ResultsBetween November 2017 and December 2018, 6747 GP FIT test requests yielded 5733 FIT results, of which 4082 (71.2%) were <4.0 μg Hb/g faeces, 579 (10.1%) were 4.0–9.9 μg Hb/g faeces, 836 (14.6%) were 10.0–149.9 μg Hb/g faeces, and 236 (4.1%) were ≥150.0 μg Hb/g faeces. The proportion of “rule out” results <4.0 μg Hb/g faeces was significantly higher than in the Getting FIT cohort (71.2% vs 60.4%, Chi squared 42.8, p < 0.0001) and the proportion of “rule in” results ≥150.0 μg Hb/g faeces was significantly lower (4.1% vs 8.1%, Chi squared 27.3,P < 0.0001).There was a 33% rise in urgent referrals across Nottingham overall during the evaluation period. 2 CRC diagnoses were made in 4082 patients who had FIT<4.0 μg Hb/g faeces. 58.4% of new CRC diagnoses associated with a positive FIT were early stage cancers (Stage I and II). The proportion of all CRC diagnoses that follow an urgent referral s rose after introduction of FIT.ConclusionsFIT allows GP's to select a more appropriate cohort for urgent investigation without a large number of missed diagnoses. FIT appears to promise a “stage migration” effect which may ultimately improve CRC outcomes.     

Faecal immunochemical testing reduces demand and improves yield of Leicester's 2-week pathway for change in bowel habit

Aim

We look at the effect of introducing the faecal immunochemical test (FIT) in the straight-to-test 2-week pathway for change in bowel habit (CIBH).

Method

The FIT in primary care triages 2-week wait (2WW) colorectal referrals for patients aged 60 years and above for straight-to-test CT colonography (CTC). We compare the impact of the FIT on numbers of 2WW CTCs, in the year before and after FIT, in both colorectal cancer (CRC) detection and cost-effectiveness at both 4 μg Hb/g faeces and 10 μg Hb/g faeces.

Results

At a threshold of 4 μg Hb/g faeces, the positive predictive value of the FIT for diagnosis of CRC is 5.0% with a negative predictive value of 99.8% and a polyp detection rate of 25.5%. The introduction of the FIT resulted in a reduction in the number of CTCs performed through the CIBH pathway from a mean of 143.9 per month prior to the FIT to 66.8 CTCs per month once the FIT was well established. Given a FIT threshold of 10 μg Hb/g the number of CTCs would be predicted to fall by 70.4% to 42.6 CTCs per month resulting in higher CRC and polyp detection rate, and an estimated annual cost saving of £238 258 in our institution.

Conclusion

The FIT use in primary care improves the yield of 2WW referrals for CIBH alone and reduces the burden and cost of investigations to exclude CRC. Improvements may be possible by increasing the cut-off employed, without adversely affecting the risk of missing a cancer.     

Changes in vitamin D metabolites during teriparatide treatment

Parathyroid hormone (PTH) increases the conversion of 25-hydroxyvitamin D [25(OH)D] to 1,25 dihydroxyvitamin D [1,25(OH)(2)D]. The purpose of this study was to assess the changes in serum concentration of vitamin D metabolites 1,25 dihydroxyvitamin D [1,25(OH)(2)D] and 25-hydroxyvitamin D [25(OH)D] during teriparatide 20 μg/day (teriparatide) therapy in the double-blind Fracture Prevention Trial of postmenopausal women with osteoporosis and in the male study of men with osteoporosis. Patients were randomized to teriparatide or placebo and received daily supplements of calcium 1000 mg and vitamin D 400-1200 IU. Serum concentrations of 1,25(OH)(2)D and 25(OH)D were measured. In women (N=336), median 1,25(OH)(2)D concentrations at 1 month increased from baseline by 27% (P<0.0001) in the teriparatide group versus -3% (P=0.87) in the placebo group (between group P<0.0001). At 12 months, the increase was 19% (P<0.0001) in the teriparatide group versus -2% (P=0.23) in the placebo group (P<0.0001). Median 25(OH)D concentrations at 12 months decreased by 19% (P<0.0001) in the teriparatide group versus 0% (P=0.13) in the placebo group (P<0.0001). In men (N=287), median 1,25(OH)(2)D concentrations at 1 month increased by 22% (P<0.0001) in the teriparatide group versus 0% (P=0.99) in the placebo group (P<0.0001). At 12 months, the increase was 14% (P<0.0001) in the teriparatide group versus 5% (P=0.004) in the placebo group (P=0.17). Median 25(OH)D concentrations at 12 months decreased by 11% (P=0.001) in the teriparatide group versus an increase of 1% (P=0.20) in the placebo group (P=0.003). Therefore, treatment with teriparatide increases 1,25(OH)(2)D concentrations and decreases 25(OH)D concentrations.     

探讨SFRP2基因甲基化在大肠癌筛查中的作用

目的:探讨粪便中分泌型卷曲相关蛋白2(secreted frizzled-related protein 2,SFRP2)基因超甲基化作为筛查大肠癌的非侵袭性工具的可行性.方法:用甲基化特异性PCR(MSP)技术分析69例大肠癌、34例腺瘤和26例增生性息肉患者的瘤组织和粪便中SFRP2基因启动子甲基化状态,30例内镜下正常的健康者作为对照.其中大肠癌患者的粪便分别于术前及术后第9天采集.同时分析SFRP2基因甲基化与肿瘤临床病理因素之间的关系.结果:SFRP2基因在91.3%(63/69)的大肠癌组织、79.4%(27/34)的腺瘤组织和53.8%(14/26)的增生性息肉组织中发生超甲基化,并在同一患者所对应的粪便中有87.0%(60/69)、61.8%(21/34)和42.3%(11/26)检测到发生超甲基化.在正常对照的结肠黏膜中没有检测到SFRP2基因甲基化,但在粪便中有2例发生了SFRP2基因甲基化.在大肠癌患者术前和术后第9天的粪便中SFRP2基因超甲基化比较差异有统计学意义(P<0.001).此外,SFRP2超甲基化与肿瘤的临床特征包括性别、年龄、肿瘤分期、位置及病理分级等无显著相关性.结论:粪便中SFRP2基因超甲基化是大肠癌的一种新的分子标志,对于非侵袭性检测大肠癌具有高度的潜力.     

细胞因子信号转导抑制因子1基因甲基化状态与胃癌幽门螺杆菌感染的关系

目的检测胃癌组织细胞因子信号转导抑制因子（SOCS-1）启动子区甲基化状态,初步探讨其甲基化状态与胃癌幽门螺杆菌（Hp）感染的关系。 方法选取2016年9月至2018年9月接受根治性手术的胃癌患者63例（胃癌组）,另选取同期行胃黏膜病理切片检查的胃黏膜息肉患者67例（对照组）,检测两组胃黏膜组织中SOCS-1基因甲基化状态,实时荧光定量PCR（RT-PCR）法及免疫印迹法检测病理组织SOCS-1 mRNA及蛋白表达水平,快速尿素酶试验检测Hp感染情况,分析胃癌组织SOCS-1甲基化与Hp感染相关性。 结果与对照组相比,胃癌组患者Hp感染阳性率升高（74.60% vs 11.94%,χ²=52.234,P<0.001）,SOCS-1启动子区CpG岛异常甲基化发生率升高（80.95% vs 5.97%,χ²=74.491,P<0.001）。Hp感染阳性胃癌患者的SOCS-1甲基化比例显著增高（95.74% vs 4.26%,χ²=26.261,P<0.001）,Hp感染阳性是SOCS-1甲基化的危险因素（OR=1.576,95% CI：1.126~2.205,P=0.035）。SOCS-1甲基化与胃癌分化程度低、TNM分期、淋巴结转移有关（χ²=11.530、9.380、11.581,均P<0.01）。与Hp感染阴性组相比,Hp感染阳性组SOCS-1 mRNA和蛋白表达量显著降低（P<0.05）。 结论Hp感染可能与SOCS-1 DNA启动子区甲基化密切相关,并通过影响SOCS-1甲基化促进胃癌发生、发展。     

长基因间非蛋白质编码RNA525在结直肠癌组织的表达及其对结直肠癌细胞增殖、侵袭的影响

目的观察长基因间非蛋白质编码RNA525(LIN00525)在结直肠癌中的表达及其对结直肠癌细胞体外增殖、侵袭的影响。方法收集2018年8月至2018年12月于复旦大学附属肿瘤医院大肠外科诊治患者的50对结肠癌及癌旁组织,通过实时定量聚合酶链式反应(Real time-PCR)检测50对样本中LINC00525的表达并分析其表达水平与临床病理参数的相关性;将LINC00525的过表达质粒转染SW480细胞,敲减质粒转染HCT-8细胞,通过细胞计数试剂盒(CCK-8)增殖实验、Transwell侵袭实验观察LINC00525对SW480及HCT-8细胞体外增殖、侵袭力的影响;通过蛋白质印迹法(Western blot)实验检测增殖相关分子增殖细胞核抗原(PCNA)及侵袭相关分子基质金属蛋白酶(MMP)-2的表达探索LINC00525调控结直肠癌细胞增殖、侵袭的分子机制;应用SPSS 20.0统计软件分析,临床资料分析使用χ²检验,两样本均数间的比较采用Student’t检验。结果50对结直肠癌组织中LINC00525的表达显著高于癌旁组织(0.721±0.028比0.204±0.016,t=15.860,P<0.01),差异有统计学意义;LINC00525的表达水平与结直肠癌淋巴转移呈正相关(χ²=10.092,P<0.01),差异有统计学意义;CCK-8实验结果显示LINC00525高表达组的吸光度(A)值显著高于对照组(2.163±0.101比1.250±0.035,t=8.525,P<0.01),差异有统计学意义;敲减LINC00525组的A值显著低于对照组(0.717±0.102比1.483±0.103,t=5.305,P<0.01),差异有统计学意义;Transwell侵袭实验结果提示过表达LINC00525组的侵袭细胞数量显著高于对照组(45.670±6.936比87.670±4.910,t=4.942,P<0.01),差异有统计学意义;敲减LINC00525组的细胞侵袭数量显著低于对照组(81.670±8.570比43.330±5.364,t=3.791,P<0.05),差异有统计学意义;Western blot显示过表达LINC00525组PCNA及MMP-2的的表达量分别高于对照组(1.72比1.00,t=5.971,P<0.01;1.58比1.00,t=7.197,P<0.01),差异有统计学意义;敲减LINC00525组PCNA及MMP-2的表达量分别低于对照组(0.39比1.00,t=7.739,P<0.01;0.33比1.00,t=12.170,P<0.01),差异有统计学意义。结论LINC00525在结直肠癌组织中高表达,LINC00525通过调控PCNA及MMP-2的表达影响结直肠癌细胞增殖和侵袭。