皮肤黑色素瘤基因突变类型及基因拷贝数变异分析

  目的  对比肢端黑色素瘤（acral melanoma, AM）与非肢端黑色素瘤（non-acral melanoma, NAM）的基因突变类型及基因拷贝数变异（copy number variation, CNV）的差异,讨论并分析其基因突变类型及拷贝数变异的意义。  方法  收集2018年1月至2021年1月新疆医科大学附属肿瘤医院收治的73例皮肤黑色素瘤患者的资料,所有患者均采用二代测序技术对46个癌症相关基因进行了全面的基因组检测。根据解剖部位分为肢端组（AMs）和非肢端组（NAMs）,比较并讨论两组患者的临床病理特征、基因突变类型及CNV的差异。  结果  两组患者在性别、年龄、肿瘤厚度、溃疡、淋巴结状态和分期等方面无显著性差异（P>0.05）。73例患者最常见的基因突变类型为BRAF突变（20.5%）、KRAS/NRAS突变（17.8%）、KIT突变（13.7%）。与AMs相比,NAMs的BRAF突变频率呈显著性差异（P<0.05）。年龄≤65岁的患者BRAF突变率与年龄>65岁的患者相比呈显著性差异（P<0.05）。此外,无溃疡的黑色素瘤比有溃疡的黑色素瘤更易出现BRAF突变（P<0.05）。两组患者中KRAS/NRAS和KIT突变率无显著性差异（P>0.05）。影响细胞周期畸变基因（CDK4/6、CCND1/2、CDKN2A）、受体酪氨酸激酶基因（EGFR、MET、ERBB2、ERBB3、PDGFRA）及抗细胞凋亡基因（BIRC2/3/5）的CNV在AMs和NAMs之间呈显著性差异（P=0.019、0.002、0.027）。  结论  通过对AMs和NAMs基因突变类型及CNV的分析,能为深入了解皮肤恶性黑色素瘤致癌模式和临床病理特征提供帮助。      

永生化上皮细胞系及肺鳞癌细胞系基因组DNA拷贝数变异的比较研究

  目的  通过分析比较支气管上皮永生化细胞系与肺鳞癌细胞系的拷贝数变异探讨肿瘤早期发展的分子机制。  方法  用人类比较基因组杂交芯片（aCGH）分别测定支气管上皮细胞系Y-BE和肺鳞癌细胞系NCI-H2170的拷贝数, 经数据校正, 对拷贝数变异基因进行GO（Gene Ontology）富集分析。  结果  永生化上皮细胞系早代Yp21仅出现了少量的DNA拷贝数变异, 而在染色体20 q11~12区段Yp21细胞与HCI-H2170细胞出现了相似的拷贝数变异结果; 永生化上皮细胞系晚代Yp113具有类神经细胞黏附基因的拷贝数增加现象; 整体来看, 从永生化上皮细胞早代到晚代, 再与肿瘤细胞比较, DNA拷贝数变异频率不断升高, 基因组稳定性逐渐下降。  结论  通过对永生化上皮细胞拷贝数变异的研究, 成功建立了肺癌癌前模型的拷贝数变异谱, 部分拷贝数变异可能代表了肿瘤发生发展早期的分子事件, 预示了细胞的潜在恶性, 这些发现可能为阐明肿瘤发生发展的分子机制提供了条件。      

乳腺浸润性导管癌17号染色体拷贝数变异与临床病理特征相关性分析

目的 乳腺癌是影响妇女健康的主要癌症之一,且发病率呈现明显上升趋势.DNA拷贝数变化(copy number variation,CNV) 能够影响基因表达,从而引起肿瘤发生,研究CNV对探索肿瘤的发病机制有很大帮助.本研究探讨乳腺癌17号染色体拷贝数变异和临床病理特征关系.方法 应用荧光原位杂交(fluorescence in situ hybridization,FISH) 法检测2010-01-01-2011-01-01新疆医科大学附属肿瘤医院乳腺润性导管癌标本283例,观察17号染色体拷贝数情况及人表皮生长因子受体2(human epidermal growth factor receptor 2,HER2) 基因扩增情况,并用免疫组织化学法检测HER2受体蛋白的表达水平.结果 283例乳腺癌中17号染色体拷贝数多倍体为43例(15.19%),双倍体为226例(79.86%),单倍体为 14例(4.95%),乳腺癌17号染色体多倍体HER2基因阳性率为53.49%(23/43).283例乳腺癌中HER2基因表达阳性为84例,阳性率为29.68%(84/283);17号染色体拷贝数不同在HER2基因是否扩增方面差异有统计学意义,χ2=9.564,P=0.007;乳腺癌17号染色体拷贝数变异在p53基因表达(χ2=8.181,P=0.017)、病理组织学分级(χ2=11.203,P=0.019)方面差异有统计学意义,与年龄、肿瘤大小和淋巴结转移等指标差异无统计学意义.17号染色体拷贝数不同组中预后存在差异,差异有统计学意义,χ2=241.363,P=0.001.结论 17号染色体拷贝数变异和乳腺癌病理特征及预后相关,检测乳腺癌中17号染色体拷贝数变异可作为指导临床治疗和判断预后的参考指标.     

新疆地区不同民族激素受体阳性乳腺癌患者CYP2D6基因多态性及拷贝数变异研究

目的:探讨新疆地区不同民族激素受体阳性乳腺癌患者CYP2D6基因多态性及拷贝数变异情况以指导临床合理用药。方法:选取2008年1月1日至2012年12月31日新疆医科大学附属肿瘤医院激素受体阳性乳腺癌患者253例,其中汉族126例、维吾尔族78例和哈萨克族49例,采用TaqMan实时荧光定量PCR技术和AccuCopyTM多重基因拷贝数检测技术进行CYP2D6基因多态性及拷贝数变异检测。结果:97.6%（247/253）的患者都检测到了CYP2D6等位基因多态性或拷贝数变异情况,最常见多态分布CYP2D6等位基因是*1、*2、*10,分布频率分别为0.577、0.320、0.462。CYP2D6的*2、*4、*10等位基因在汉族、维吾尔族及哈萨克族乳腺癌患者中分布频率分别为（0.023 8、0.038 4、0.042 9）、（0.031、0.038、0.163）、（0.635、0.372、0.184）,差异有统计学意义。汉族、维吾尔族、哈萨克族乳腺癌患者CYP2D6代谢表型及拷贝数情况分布差异无统计学意义。结论:在新疆地区汉族、维吾尔族、哈萨克族激素受体阳性乳腺癌患者CYP2D6基因在代谢表型多态性及拷贝数变异情况方面未发现明显的民族差异性,但*2、*4、*10等位基因在不同民族间分布有差异,为他莫昔芬个体化治疗的进一步研究提供了一定依据。     

肝癌组织中MPDZ基因拷贝数和表达变化及其诊断预后价值分析

目的：探讨MPDZ基因在肝癌组织中的拷贝数和表达变化情况,并评价其与临床病理指标之间的关系以及对肝癌诊断和预后判断的价值。方法：利用TCGA数据库分析MPDZ基因在299例肝癌组织中的拷贝数改变情况及其与表达的关系;同时分析TCGA数据中MPDZ基因在282例肝癌组织和50例癌旁组织中的表达及其与临床病理指标和生存预后的关系。结果：MPDZ基因在肝癌组织中存在明显的拷贝数变异,且拷贝数变异的主要类型为拷贝数缺失（31.44%）;同时MPDZ基因拷贝数缺失后MPDZ mRNA表达下调。MPDZ基因在肝癌组织中的表达显著低于癌旁组织,差异具有统计学意义（P < 0.05）。Kaplan-Meier分析显示,MPDZ基因低表达患者的生存时间短于高表达患者,差异具有统计学意义（Log-rank=12.48,P < 0.05）。受试者工作特征曲线（ROC）分析结果表明,MPDZ基因表达是一个灵敏度和特异度较好的肝癌诊断指标（AUC=0.86）。结论：MPDZ基因拷贝数缺失可能与肝癌的发生有关,MPDZ基因的表达对肝癌的诊断及生存预后判断具有参考价值。     

乳腺癌的基因分型及基底细胞样型乳腺癌

0 引言 乳腺癌具有很大的分子异质性,传统乳腺癌分类以形态学为基础,它不能很好的反映肿瘤发生的分子机制和判断预后.Perou等[1]首先依据基因表达类型对乳腺癌进行分型,并将一组具有肌上皮/基底细胞基因表达及免疫表型特征的乳腺癌命名为基底细胞样型乳腺癌(basal-like breast carcinoma,BLBC).本文结合近年来的文献,对乳腺癌基因分型的建立及BLBC的临床病理特征作一综述.     

基因分型对乳腺癌诊治和预后判断的指导作用

乳腺癌是女性的常见恶性肿瘤之一,其临床治疗主要是以经典的TNM和临床分期为指导。近年来兴起的对乳腺癌相关的分子遗传学研究取得了令人瞩目的进展,提出了乳腺癌基因分型这一新概念。基因诊断对乳腺癌的辅助化疗以及预后的判断相对传统的病理分型有更高的专一性,为乳腺癌的治疗起到了更好的指导作用,本文主要对基因分型进行介绍并总结了几种与乳腺癌预后相关的基因标志,探讨其在临床治疗中的实际应用。     

LKB1基因在不同基因亚型乳腺癌中表达的意义

目的:检测LKB1基因在不同基因亚型乳腺癌中的表达,并探讨其在乳腺癌中表达的意义.方法: 应用组织芯片及免疫组织化学法对180例乳腺浸润性导管癌、18例导管原位癌及20例正常乳腺组织中LKB1基因、 ER 、PR、HER-2、CK56、和P53 的表达进行检测;根据最近提出的乳腺癌分子分型进行分组,对LKB1基因在不同基因亚型中的表达及与其他临床病理特征的关系进行统计学分析. 结果: LKB1基因在乳腺导管原位癌的阳性率为66.11%(11/18),浸润性导管癌的阳性率为36.67%(66/180),两组间差异有统计学意义(P<0.05);在浸润性导管癌中,LKB1基因在淋巴结转移组阳性率为47.61%(40/84),无转移组阳性率为83.33%(80/96),两组间差异具有统计学意义(P<0.01);在不同基因亚型中中LKB1基因的表达差异具有统计学意义(P<0.05);其中Luminal A组与HER-2+组及三阴性组两两比较差异具有统计学意义(P<0.05).结论: LKB1基因的在乳腺浸润性导管癌中的表达明显低于导管原位癌,与淋巴结转移呈负相关,且在生物学行为更为恶性的HER-2+组和三阴性组具有更低的表达水平,提示LKB1基因对乳腺癌的浸润与转移过程有抑制作用.     

全基因组拷贝数变异评分在肺腺癌诊断及预后预测中的价值

目的探讨全基因组拷贝数变异(WGCNV)检测和评分系统在肺腺癌诊断和预后预测中的价值。方法应用显微微切割技术获取76例肺腺癌患者肿瘤组织标本91份和癌旁正常对照组织样本20份, 通过全基因组扩增(WGA)富集DNA并构建测序文库, 进行二代测序。评估肺腺癌手术和恶性胸水细胞学标本的肿瘤细胞核级别改变, 在肿瘤核细胞大小、核分裂象计数、细胞核不典型性和不典型核分裂4个方面进行分级。评价WGCNV评分的预测预后的价值, 根据受试者工作特征(ROC)曲线分析WGCNV评分在肺腺癌中的诊断价值。结果 20例癌旁正常肺组织样品WGCNV改变作为正常对照, 所有肿瘤标本WGCNV评分为0～9.95分, 中位WGCNV评分为2.7分。核级别评分与WGCNV评分之间存在正相关(r=0.780 90, P<0.000 1)。中评分组(1.74～4.23分)相对低评分组(<1.74分)的风险比为4.11(95%CI为0.72～23.57), 高评分组(>4.23分)相对低评分组的风险比为2.07(95%CI为0.30～14.12), 中、高评分组风险呈上升趋势。ROC曲线分析显示, 当临...     

Ultrasonographic Features of Triple-Negative Breast Cancer: a Comparison with Other Breast Cancer Subtypes

Background: Triple-negative breast cancer (TNBC) is known to be associated with aggressive biologic featuresand a poor clinical outcome. Therefore, early detection of TNBC without missed diagnosis is a requirementto improve prognosis. Preoperative ultrasound features of TNBC may potentially assist in early diagnosis ascharacteristics of disease. Purpose: To retrospectively evaluate the sonographic features of TNBC compared toER (+) cancers which include HER(-) and HER2 (+), and HER2 (+) cancers which are ER (-). Materials andMethods: From June 2012 through June 2014, sonographic features of 321 surgically confirmed ER (+) cancers(n=214), HER2 (+) cancers (n=66), and TNBC (n=41) were retrospectively reviewed by two ultrasound specialistsin consensus. The preoperative ultrasound and clinicopathological features were compared between the threesubtypes. In addition, all cases were analyzed using morphologic criteria of the ACR BI-RADS lexicon. Results:Ultrasonographically, TNBC presented as microlobulated nodules without microcalcification (p=0.034). A lowerincidence of ductal carcinoma in situ (p<0.001), invasive tumor size that is>2 cm (p=0.011) and BI-RADS category4 (p<0.001) were significantly associated with TNBC. With regard to morphologic features of 41 TNBC cases,ultrasonographically were most likely to be masses with irregular (70.7%) microlobulated shape (48.8%), becircumscribed (17.1%) or have indistinct margins (17.1%) and parallel orientation (68.9%). Especially TNBCmicrolobulated mass margins were more more frequent than with ER (+) (2.0%) and HER2 (+) (4.8%) cancers.Conclusions: TNBC have specific characteristic in sonograms. Ultrasonography may be useful to avoid misseddiagnosis and false-negative cases of TNBC.     

Comparison of Time-Varying Pattern of Recurrence in Chinese Breast Cancer Patients with Different Molecular Subtypes: A Single-Center Retrospective Study

Background: To compare the time-varying recurrence patterns of different molecular subtypes of breast cancer inthe contemporary era with those in the past era. Patients and Methods: This retrospective study included14627 consecutive invasive breast cancer patients who underwent surgery from 2008 to 2016 at Fudan UniversityShanghai Cancer Center. We defined the period from 2013 to 2016 as the contemporary era and that from 2008 to2012 as the past era. Five subtypes were defined according to the immunohistochemistry results. Emphasis wasmade on the changing patterns of recurrence for patients with different molecular subtypes changed between thetwo treatment eras. Kaplan–Meier survival curves were generated, and the hazard function was used to estimatethe annual recurrence hazard. Results: By the end of follow-up (median, 68.1 months), 1429 patients (9.77%)experienced recurrence and metastasis. The annual recurrence risks of the entire population and each moleculartype in 2013–2016 were reduced compared with those in 2008–2012. Luminal A and triple-negative patients in2013–2016 showed a significantly lower recurrence hazard curve than those in 2008–2012. The recurrence hazardcurve of luminal B (HER2−) and HER2+ subtypes was lower in 2013–2016 than in 2008–2012. The recurrencerisk of the luminal B (HER2+) subtype was reduced substantially during 2013–2016, showing a delay of two years.Conclusions: With early detection of disease recurrence and improved treatment strategies, the recurrence patterns of different molecular subtypes of breast cancer have changed. The time-varying recurrence patternsobserved during the contemporary era are significant for treatment decision-making and clinical trial planning.     

Age‐Specific Changes in Intrinsic Breast Cancer Subtypes: A Focus on Older Women

Purpose.

Breast cancer (BC) is a disease of aging and the number of older BC patients in the U.S. is rising. Immunohistochemical data show that with increasing age, the incidence of hormone receptor-positive tumors increases, whereas the incidence of triple-negative tumors decreases. Few data exist on the frequency of molecular subtypes in older women. Here, we characterize the incidence and outcomes of BC patients by molecular subtypes and age.

Patients and Methods.

Data from 3,947 patients were pooled from publicly available clinical and gene expression microarray data sets. The PAM50 algorithm was used to classify tumors into five BC intrinsic subtypes: luminal A, luminal B, HER2-enriched, basal-like, and normal-like. The association of age and subtype with recurrence-free survival (RFS), overall survival, and disease-specific survival (DSS) was assessed.

Results.

The incidence of luminal (A, B, and A+B) tumors increased with age (p < .01, p < .0001, and p < .0001, respectively), whereas the percentage of basal-like tumors decreased (p < .0001). Among patients 70 years and older, luminal B, HER2-enriched, and basal-like tumors were found at a frequency of 32%, 11%, and 9%, respectively. In older women, luminal subtypes had better outcomes than basal-like and HER2-enriched subtypes. After controlling for subtype, treatment, tumor size, nodal status, and grade, increasing age had no impact on RFS or DSS.

Conclusion.

More favorable BC subtypes increase with age, but older patients still have a substantial percentage of high-risk tumor subtypes. After accounting for tumor subtypes, age at diagnosis is not an independent prognostic factor for outcome.     

DNA Repair Gene Patterns as Prognostic and Predictive Factors in Molecular Breast Cancer Subtypes

DNA repair pathways can enable tumor cells to survive DNA damage induced by chemotherapy and thus provide prognostic and/or predictive value. We evaluated Affymetrix gene expression profiles for 145 DNA repair genes in untreated breast cancer (BC) patients (n = 684) and BC patients treated with regimens containing neoadjuvant taxane/anthracycline (n = 294) or anthracycline (n = 210). We independently assessed estrogen receptor (ER)‐positive/HER2‐negative, HER2‐positive, and ER‐negative/HER2‐negative subgroups for differential expression, bimodal distribution, and the prognostic and predictive value of DNA repair gene expression. Twenty‐two genes were consistently overexpressed in ER‐negative tumors, and five genes were overexpressed in ER‐positive tumors, but no differences in expression were associated with HER2 status. In ER‐positive/HER2‐negative tumors, the expression of nine genes (BUB1, FANCI, MNAT1, PARP2, PCNA, POLQ, RPA3, TOP2A, and UBE2V2) was associated with poor prognosis, and the expression of one gene (ATM) was associated with good prognosis. Furthermore, the prognostic value of specific genes did not correlate with proliferation. A few genes were associated with chemotherapy response in BC subtypes and treatment‐specific manner. In ER‐negative/HER2‐negative tumors, the MSH2, MSH6, and FAN1 (previously MTMR15) genes were associated with pathological complete response and residual invasive cancer in taxane/anthracycline‐treated patients. Conversely, PMS2 expression was associated with residual invasive cancer in treatments using anthracycline as a single agent. In HER2‐positive tumors, TOP2A was associated with patient response to anthracyclines but not to taxane/anthracycline regimens. In genes expressed in a bimodal fashion, RECQL4 was significantly associated with clinical outcome. In vitro studies showed that defects in RECQL4 impair homologous recombination, sensitizing BC cells to DNA‐damaging agents.