Effectiveness of an adriamycin immunoconjugate that recognizes the c-erbB-2 product on breast cancer cell lines

Adriamycin (ADM) was chemically conjugated to a murine monoclonal antibody, A0011, which recognizes the c-erbB-2 product, via a disulfide bond using N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP) and 2-iminothiolane (2-IT). The molar ratio of ADM to the monoclonal antibody ranged from 15:1 to 25:1 and enzyme-linked immunosorbent assay (ELISA) showed that the binding activity of the conjugate was almost retained. We compared the efficacy of A0011 alone, ADM alone, the A0011-ADM conjugate, and a nonspecific murine IgM-ADM conjugate, against the human breast cancer cell lines SK-BR-3, MDA-MB-361, MCF-7, and BT-20. The A0011-ADM conjugate was observed to be ten times more cytotoxic to the cell lines overexpressing the c-erbB-2 product, namely, SK-BR-3 and MDA-MB-361, than free ADM, but it showed weak cytotoxicity against the cell lines with a low level of c-erbB-2 product expression, namely, MCF-7 and BT-20. However, free A0011 and nonspecific murine IgM-ADM conjugate showed no cytotoxicity toward any of the four cell lines, while the addition of a tenfold molar excess of A0011 inhibited conjugate cytotoxicity. These data suggest that conjugate cytotoxicity is antibody-mediated. Moreover, conjugate cytotoxicity at 10^–6M was correlated with antigen volume, and the data were fitted to the regression equation y=–11.63logX+116.38 where the correlation coefficient =0.950. Our results indicate that targeting therapy aiming at the c-erbB-2 product may be useful in the treatment of breast cancers overexpressing the c-erbB-2 product.     

Immunohistochemical overexpression of C-erbB-2 in the prognosis of breast cancer

Immunohistochemical c-erbB-2 protein overexpression was detected in 34 of 124 (27.4%) paraffin-embedded breast cancer specimens. Although no difference was seen between the c-erbB-2 positive and negative groups in 5-year disease-free survival, 5-year overall survival was significantly less favorable in the c-erbB-2 positive group. Furthermore, patients graded as having positive c-erbB-2 staining and aneuploid DNA showed significantly poorer survival than those in other categories. The significant prognostic factors, determined by a multivariate analysis, were nodal status and c-erbB-2 overexpression. Our findings therefore suggest that c-erbB-2 expression is a prognostic factor in breast cancer and that it could be useful in the determination of postoperative adjuvant therapy.     

The role of c-erbB-2/HER2/neu in breast cancer progression and metastasis

Gene amplification and/or overexpression of the c-erbB-2/HER2/neu tyrosine kinase are linked with poor prognosis in breast cancer. This is manifest in shorter disease-free intervals, increased risk of metastasis, and resistance to many types of therapy. The molecular mechanisms and signaling circuitry underlying these phenomena are now being elucidated. c-erbB-2, although having no known soluble ligand, is transactivated by heterodimerization with other family members (EGFR, c-erbB-3, c-erbB-4). Receptor activation potentiates tumor cell motility, protease secretion and invasion, and also modulates cell cycle checkpoint function, DNA repair, and apoptotic responses. Since it is expressed at low levels in normal adult tissues, c-erbB-2 is an ideal target for therapy. There is reason for optimism that agents targeting c-erbB-2 signaling will have profound and selective effects in breast cancer, either as single agents or more likely in combination with other therapeutic agents, to enhance their potency.     

p53、c-erbB-2和组织蛋白酶D与腋窝淋巴结阴性乳腺癌患者预后的关系

目的：探讨p53,c-erbB-2和组织蛋白酶D（cathepsinD)在评估腋窝淋巴结阴性乳腺癌（NNBC）患者预后中的价值,方法：用免疫组织化方法,检测110例T1－2N0M0期乳腺癌患者原发灶癌组织中p53,c-erbB-2和cathepsinD水平,并用单因素和多因素统计方法对其结果与肿瘤的临床特征有患者预后之间的关系进行了分析,结果：本组肿瘤直径＞3cm的患者比＜3cm 患者的远处转移率高,无瘤生存率和总生存率下降,差异有显著性意义（分别P＜0．03,P＜0．01,P＜0．05）,cathepsin阴性患者的肿瘤远处转移率,无瘤生存率,分别为8．25％,82．61％,阳性患者为33．06%,57．41％,差异均有极显著性意义（P＜0．01）,c-erbB-2 阴性患者及p53表达状态与患者的肿瘤远处转移率,无瘤生存率和总生存率无关,62例未作全身治疗的患者,cathepsin阴性患者的肿瘤远处转移率,无瘤生存率、分别为8．92％,77．04％,阳性患者为44．61％,43．46％,差异均有极显著性意义（P＜0．01）,c-erbB-2阴性患者的肿瘤远处转移率,无瘤生存率,和总生存率分别为16．70％,70．11％,88．30％,阳性患者为41．26％,44．44％,68．65％,差异均有显著性意义（P＜0．05）,53表达状态与患者的肿瘤远处转移率,无瘤生存率和总生存率无关,多因素分析表明肿瘤大小和cethepsinD表达水平与患者的远处转移率,无瘤生存率和总生存率无关,多因素分析表明肿瘤大小和cathepsinD表达水平与患者的远处转移率,无瘤生存率无关（P＜0．05）,48例接受全身治疗的患者p53,c-erbB-2及cathepsinD的表达状态与患者预后无关,结论：肿瘤大小和cathepsinD 水平可作为评估腋窝淋巴结阴性乳腺癌患者预后的独立指标,对患者治疗方案的选择具有一定的指导意义。     

Higher expression of oncoproteins c-myc,c-erbB-2/neu,PCNA, and p53 in metastasizing colorectal cancer than in nonmetastasizing tumors

Background: Expression of individual oncogenes may predict outcome in patients with metastatic colorectal cancer (CRC). We studied the oncogene profile in the tumors of patients with CRC and assessed their value as predictors of liver metastases. Methods: The oncoproteins c-myc, c-erbB-2/neu (c-neu), PCNA and p53, were measured by immunohistochemistry in sections of metastasizing human CRC (n=34) and their liver secondaries as well as in sections of nonmetastasizing CRC (n=25). Results: The metastasizing primary CRC expressed proliferating-cell nuclear antigen (PCNA), c-neu, and c-myc at significantly higher levels than the nonmetastasizing primary cancer. p53 was also overexpressed in the metastatic group compared with the nonmetastasizing CRC, but this difference was not significant. The frequency of expression of all these markers was similar in the metastasizing primary CRC and the liver secondaries from the same patients. There was no correlation between the expression of the individual markers and histological grade, DNA ploidy, and subsequent local recurrence and lung metastasis and survival. However, when both groups were assessed together, positive expression of c-myc was more likely to occur in poorly differentiated tumors, whereas PCNA expression increased with more advanced Dukes stages. Conclusion: These results suggest that the overexpression of c-myc, c-neu, PCNA, and p53 may occur in CRC that are likely to metastasise to the liver.     

No effect of timing of biopsy in the menstrual cycle on incidence of bone marrow micrometastasis in patients with breast cancer

Background: The timing of breast cancer excision relative to the menstrual cycle has been debated to be of significant prognostic value. The differences in survival relative to the timing in the menstrual cycle have been attributed to the incidence of micrometastasis. Methods: All patients underwent bone marrow aspiration after the diagnostic surgical biopsy, immediately before definitive surgery. The timing of the diagnostic surgical biopsy in the menstrual cycle was calculated according to Senie. Monoclonal antibodies to epithelium-specific antigens were used to detect bone marrow micrometastasis (BMM). Results: This study reports on the effect of the phase of the menstrual cycle on incidence of BMM after surgical biopsy. Ninety-two patients with regular cycles underwent bone marrow aspiration an average of 12 days (range 0–32 days) after biopsy. Thirty-nine patients had undergone biopsy during the follicular phase and 53 patients during the luteal phase. BMM were detected in 31% of patients (29 of 92). We observed BMM in 33% of patients (13 of 39) in the follicular phase and in 30% of patients (16 of 53) in the luteal phase. This difference is not significant (p>0.70). The mean number of cells detected and the presence of clumps of cells is similar in the two groups. Conclusions: The incidence and characteristics of bone marrow micrometastases are independent of the timing of diagnostic excision biopsy in the menstrual cycle. Dr. Singhal is now with the Department of Surgery, University of Wales College of Medicine, Cardiff, CF4 4XN United Kingdom.     

Characterization of cell surface antigens expressed in the HMA-1 breast cancer cell line

This report describes the characterization of an estrogen receptor-positive breast cancer cell line, HMA-1, established from a breast cancer patient, based on the expression of tumor-associated antigens (TAAs), the HLA-DR antigen, and the c-erbB-2 proto-oncogene product. In flow cytometric and immunohistochemical analyses, HMA-1 was found to express increased levels of several TAAs including MUC1, TAG-72 (sialyl Tn), Tn, T, sialyl Le^a, Le^x, and Le^y, HMA-1 also expressed enhanced levels of the HLA-DR antigen and c-erbB-2 protein. These results indicate that HMA-1 is a unique cell line with abundant TAAs which may serve as an appropriate breast cancer cell line for application in the multidisciplinary research of breast cancer.     

骨桥蛋白在人正常月经周期子宫内膜的表达

目的研究骨桥蛋白(osteopontin,OPN)在人正常月经周期子宫内膜的表达。方法采用免疫组织化学法对51例人正常月经周期子宫内膜OPN蛋白质进行定位和定量检测,Western blotting法测定增殖期及分泌期OPN蛋白水平,采用逆转录-聚合酶链反应(RT-PCR)方法检测18例人正常月经周期子宫内膜增殖期及分泌期OPN mRNA的表达。结果OPN蛋白质主要分布在正常子宫内膜腺上皮细胞,增殖早期、中期无表达,增殖晚期出现,分泌中期、晚期表达最强。月经期子宫内膜腺上皮细胞表达较强。OPN mRNA表达分泌期强于增殖期(P<0.05)。Western blotting检测结果显示正常子宫内膜组织中存在OPN蛋白,分泌期含量较高。结论OPN蛋白质及mRNA在正常子宫内膜中的表达呈现明显周期依赖性。分泌中、晚期表达增强提示其可能参与胚胎着床。     

不同致密度乳腺腺体的表观弥散系数与月经周期的关系

目的探讨不同致密度正常乳腺腺体的ADC值与月经周期的关系。方法将50名(100侧乳腺)女性志愿者按腺体致密度分为致密型(40侧)、中间型(36侧)、脂肪型(24侧),分别在月经期、增殖期及分泌期行T2WI-TIRM、DWI检查,并对各期乳腺腺体的ADC值进行测量,采用单因素方差分析、配对t检验比较各月经周期中不同腺体类型的ADC值及同一腺体类型在不同月经周期ADC值的差异。结果月经期时致密型、中间型及脂肪型腺体的ADC值分别为(2.08±0.07)×10-3 mm2/s、(2.09±0.09)×10-3 mm2/s、(1.90±0.35)×10-3 mm2/s;增殖期时各类腺体的ADC值分别为(1.97±0.16)×10-3 mm2/s、(2.00±0.17)×10-3 mm2/s、(1.81±0.54)×10-3 mm2/s;分泌期时各类腺体的ADC值分别为(2.01±0.08)×10-3 mm2/s、(2.02±0.05)×10-3 mm2/s、(1.82±0.21)×10-3 mm2/s;各腺体类型在月经期ADC值与增殖期及分泌期差异均有统计学意义(P均0.05),各月经周期脂肪型腺体的ADC值与致密型及中间型腺体ADC值差异均有统计学意义(P均0.01)。结论进行乳腺DWI检查时应考虑腺体致密度及月经周期对乳腺的影响,尽量选择月经结束后1周内完成。     

NOEY2 mutations in primary breast cancers and breast hyperplasia

PurposeThe NOEY2 gene mutations and protein expression in human breast cancers, adjacent breast tissues and breast benign lesions were analysed to explore the potential correlation between the mutation spectrum and breast cancer development and progression.Experimental designThe promoter, exon and intron regions of NOEY2 gene were amplified by polymerase chain reaction (PCR) with DNA extracted from 50 human breast cancer and corresponding adjacent breast tissues as well as 50 breast benign lesions, respectively. The PCR products were then sequenced and analysed. The effect of mutations on the expression of NOEY2 protein by immunohistochemistry were proven as well.ResultsTwenty-one of 50 (42%) breast cancer mutations were identified in promoter (11 cases) and exon 2 (seven cases on untranslation region and three on coding region) and 17 of 50 (34%) adjacent breast tissues (all were atypical hyperplasia lesions) occurred mutations, including six promoter mutations and 11 exon 2 changes (10 cases on untranslation region and one on coding region). Interestingly, the mutations were identified in both breast cancers and the corresponding adjacent breast tissues collected from the same patient in seven of them. No mutation was identified in all benign breast tissues. Immunohistochemical analysis showed that two of 17 mutational adjacent breast tissue samples were NOEY2 immunoreaction negative, and in all 21 mutations of breast cancers five cases were of loss of NOEY2 expression. All mutations with immunoreaction negative factor were located at promoter and/or exon 2 coding region. NOEY2 gene mutations were not correlated with patient ages, histological types, tumour sizes, histological grades, clinical stages, axillary lymph node metastases or with the condition of hormone receptor (ER, PR) expression and HER2 amplification.ConclusionsThe mutations of human NOEY2 were identified in human breast cancers and the corresponding adjacent breast tissues. The hot mutation spots were its promoter and exon 2 regions, and those occurring at the exon2 coding region and part of the promoter may alter the expression of NOEY2. The presence of NOEY2 mutations in human breast cancer and early-stage lesions indicates that NOEY2 mutations may be partly associated with breast tumourigenesis.     

Loss of c-met protooncogene in primary and metastatic sites of breast cancer

Background: It has been proposed that clones of tumor cells acquire higher metastatic potential as a result of specific genetic alterations. This study was designed to determine the role of the c-met protooncogene in systemic spread by comparing the loss of the c-met protooncogene between primary and metastatic breast carcinomas. Methods: Only patients who had not received chemotherapy or radiotherapy in the preceding 6 months were included in this study. Histologically proven malignant tissue was obtained from the primary tumor, involved nodes, and distant metastatic and recurrent tumors of patients with breast carcinomas. Allelic loss of the c-met protooncogene in tumor tissue was determined by Southern blotting using a polymerase chain reaction-generated 347-bp human met-H probe. Restriction digestion was performed usingTaq I andMsp I, with the patient's lymphocyte DNA as controls. Results: Of 52 patients, lymphocyte DNA from 36 patients was heterozygous for the c-met protooncogene (69% informative). Forty-six tumors from these 36 patients were analyzed. Four of 30 primary tumors (13%) showed allelic loss of c-met. Of the nine nodal metastases examined, three (33%) showed allelic loss of the c-met protooncogene. Of seven distant metastatic breast tumors or recurrent disease, two (29%) showed allelic loss (both in patients with skin metastasis in the chest wall). Conclusions: Allelic loss of the c-met protooncogene was detected in both primary (13%) and metastatic sites (31%) of breast cancer. Although a higher proportion of allelic loss of c-met was noted in nodal and distant/recurrent disease, the difference when compared with the primary tumor was not statistically significant. These findings indicate a limited role of the c-met protooncogene in breast cancer metastases.     

月经周期中内源性大麻素与性激素和促性腺激素的相关性研究分析

目的分析月经周期中不同时期正常女性血浆中N-花生四烯酸氨基乙醇(AEA)的水平变化及其与性激素和促性腺激素的相关性。方法选择2015年5~10月因输卵管梗阻或男方因素到本院生殖医学科就诊并欲行IVF助孕、月经周期和排卵正常的育龄妇女为研究对象。根据实验目的分为横断面研究组(79例)和纵向研究组(10例),检测AEA在两组女性月经周期中不同时期的变化,以及AEA与FSH、LH、E2、P之间的相关性。结果横断面研究组中早卵泡期、晚卵泡期、排卵期和黄体期AEA水平分别为(9.71±0.86)ng/ml、(10.61±1.05)ng/ml、(12.24±0.73)ng/ml和(7.46±0.71)ng/ml。排卵期AEA水平最高,黄体期最低(P0.05)。纵向研究组4个时期AEA水平分别(8.76±0.91)ng/ml、(11.61±1.28)ng/ml、(13.85±1.18)ng/ml、(8.50±1.08)ng/ml,排卵期AEA水平最高,黄体期最低(P0.05)。横断面研究组和纵向研究组各个时期的AEA水平比较均无显著性差异(P0.05)。AEA与FSH、LH、E2之间存在显著的正相关(P0.05),与P水平则无显著相关性(P=0.067)。结论正常女性血浆内AEA的浓度随月经周期的变化而变化,且与性激素和促性腺激素呈明显相关。但AEA与FSH、LH、E2、P之间的具体作用机制,有待进一步研究。     

组织蛋白D和E-钙黏附蛋白与乳腺癌患者预后的关系

目的探讨E-Ca、Ca-D在乳腺癌预后中的作用及相互关系。方法对106例浸润性乳腺癌患者的肿瘤组织石蜡切片采用免疫组织化学染色技术标记E-Ca、Ca-D。结果肿瘤间质内Ca-D表达强度与复发呈正相关。E-Ca表达在导管癌内表达强于小叶癌。乳腺癌组织中nm23-H1蛋白表达明显降低,与生存期呈正相关,与复发期呈负相关（P〈0.05）。MVD在复发组高于对照组。结论①Ca-D、MVD、nm23-H1在乳腺癌组织内共同参与癌组织的浸润和转移,可作为预后的判断指标。②E-Ca作为预后判断不理想,但可以作为差分化小叶癌和导管癌的鉴别诊断依据。     

乳腺癌c-erbB-2与雌孕激素受体及淋巴结转移的相关性分析

目的：分析乳腺癌c-erbB-2的表达与ER、PR，以及腋窝淋巴结、肿瘤大小、病理类型的相关性。方法：收集术前未行新辅助化疗而行手术切除加腋窝清扫的乳腺癌病人346例，采用免疫组化法检测c-erbB-2、ER及PR；比较c-erbB-2表达与ER、PR、腋窝淋巴结、肿瘤大小以及病理类型之间的关系。结果：c-erbB-2阳性表达者占48．2％，腋窝淋巴结有转移者的c-erbB-2阳性表达为69．6％，而淋巴结阴性者则为34．6％（P〈0．001）。ER、PR均阳性且c-erbB-2表达也阳性者为38．1％；而ER、PR均阴性、c-erbB-2阳性者达64．6％（P〈0．001）。仅ER阳性和阴性者的c-erbB-2阳性表达分别是40．7％和69.2％（P（0．001）。仅PR阳性和阴性者的c-erbB-2阳性分别为46．4％和49．7％（P〉0．05）。〈2cm和2～5cm肿瘤的c-erbB-2阳性率分别是47．1％和48．8％（P〉0．05）。导管内癌和黏液癌的c-erbB-2表达阳性率分别达73．8％和60．0％。结论：c-erbB-2阳性表达与腋窝淋巴结的转移呈正相关，与ER呈负相关，与PR和肿瘤大小不相关。c-erbB-2是判断乳腺癌预后的良好指标。     

乳腺癌广泛导管内癌成分与组织蛋白酶D Ki-67及单链DNA表达的关系及临床意义

目的 探讨乳腺癌广泛导管内癌成分（extensive intraductal component，EIC）、粉刺状坏死（comedo necrosls）、组织学分级与组织蛋白酶D（eathepsin D）、Kiab7、单链DNA（ssDNA）及c-erbB-2表达的关系。方法检测日本德岛乳腺门诊与东德岛医院1996年1月至1997年4月连续手术切除的59例乳腺浸润性导管癌标本。用免疫组化方法，检测59例乳腺浸润性导管癌中组织蛋白酶D（eathepsin D）、Ki-67、单链DNA及c-erbB-2的表达。结果EIC阳性组，肿瘤上皮细胞中的组织蛋白酶D为高表达（P=0．021），肿瘤间质细胞中的组织蛋白酶D表达差异无显著性意义。粉刺状坏死的出现，与c-erbB-2癌基因、肿瘤上皮细胞中的组织蛋白酶D和细胞增殖指数相关（P分别为0．042，0．006及0．013）。结论肿瘤上皮细胞中的组织蛋白酶D可能参与了EIC与粉刺状坏死的形成。EIC与粉刺状坏死的检测有临床应用价值。     

Decreased expression of the DCC gene in human breast carcinoma

Inactivation of the deleted in colon cancer (DCC) gene on chromosome 18 is known to be associated with the tumorigenesis and metastasis of colorectal cancer. In the present study, we investigated the expression of DCC and the c-erbB-2 product in surgical specimens from 45 patients with breast cancer by immunohistochemical staining, and found the expression of DCC to be decreased in 23 (51%) tumors. In 8 years of follow-up, 11 of 22 (50%) patients with DCC-positive staining tumors, and 17 of 23 (74%) patients with DCC-negative tumors developed recurrence. The stratified analysis, according to the status of axillary lymph node metastasis, showed the same tendency. Overexpression of erbB-2 was detected in 13 (29%) of the 45 breast cancer specimens, but there were no differences in the relapse rate between patients with erbB-2 positive and those with erbB-2 negative tumors. Although the individual alteration of DCC or erbB-2 did not possess independent prognostic significance for the prediction of recurrence, patients with tumors having the double alteration of DCC-negative and erbB-2-positive showed adverse relapse-free survival (0.025<P<0.05). These findings suggest that a decrease in DCC expression and erbB-2 overexpression may influence the progression of breast carcinoma.     

Correlation between the expression of oncogenes ras and c-erbB-2 and the biological behavior of bladder tumors

Summary Expression of the ras and the c-erbB-2 oncogene products was investigated in 56 cases of human bladder transitional cell carcinoma and 6 samples of human normal bladder tissue using an immunohisto-chemical method. Thirty of the 56 cases of bladder tumor were found to be immunohistologically positive with the monoclonal anti-ras p21 antibody, while 19 of 56 cases were positive with the polyclonal anti-c-erbB-2 oncoprotein antibody. All 6 controls were negative with both antibodies. The ras p21 positive staining was found more frequently in the well or moderately differentiated, superficial and non-recurrent tumors than in the poorly differentiated (p<0.01), muscle invasive (p<0.05) and recurrent tumors (p<0.01), while the c-erbB-2 gene product was more commonly detected in high-grade (p<0.01), invasive (p<0.01) and recurrent tumors (p<0.05). Thus, the expression of either ras or c-erbB-2 was closely associated with the histological grade, clinical stage and recurrence of bladder transitional cell carcinomas.     

不同浓度罗哌卡因对乳腺癌MCF-7细胞增殖和细胞周期的作用研究

目的观察不同浓度罗哌卡因对乳腺癌MCF-7细胞增殖和细胞周期的影响并探讨其机制。方法人乳腺癌细胞MCF-7接种于培养板培养24h,随机分为四组:对照组(C组)、罗哌卡因100μg/ml组(R1组)、罗哌卡因200μg/ml组(R2组)和罗哌卡因400μg/ml组(R3组)处理乳腺癌MCF-7细胞48h后,检测其细胞增殖能力(细胞活力)和细胞周期。检测R3组作用于MCF-7细胞48h后TCF-4、beta-catenin的蛋白表达水平。结果 R2、R3组MCF-7细胞活力明显低于C组(P0.05);R1、R2、R3组MCF-7细胞G0/G1期细胞明显少于C组,S期和G2/M期细胞明显多于C组(P0.05);R3组TCF-4和beta-catenin蛋白表达水平明显低于C组(P0.05)。结论罗哌卡因可能通过下调TCF-4和beta-catenin蛋白表达水平抑制人乳腺癌MCF-7细胞增殖。     

Role of cathepsin D in prostatic cancer cell growth and its regulation by brefeldin A

 We investigated a possible relationship between brefeldin A (BFA), an antibiotic, and cathepsin D (Cat.D), a lysosomal protease, in prostate cancer proliferation. Effects of BFA (30 ng/ml) were examined on the growth of three human prostatic cancer cell lines, PC-3, DU-145, and LNCaP cells. Its effect on Cat.D in these cancer cells was assessed by Western blots and compared with Cat.D expressed in clinical prostate specimens (n=55). BFA profoundly (>70%) inhibited the growth of all three cancer cell lines. Western blots revealed that expression of procathepsin D (Pro.Cat.D) was markedly increased with BFA, whereas actively proliferating (control) cells greatly exhibited mature Cat.D. Analysis of prostate specimens then showed predominant Pro.Cat.D expression in non-cancerous tissues while also showing enhanced expression of mature Cat.D in all cancer specimens. Therefore, BFA-induced growth inhibition in prostatic cancer cells is associated with a blocking of Cat.D maturation (activation), suggesting a possible role of Cat.D in prostate cancer proliferation/development.     

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。