Relationship between CYP1A1 polymorphisms and invasion and metastasis of breast cancer

Objective:To investigate the relationship between CYPIA1 genetic polymorphisms and the invasion and metastasis of breast cancer.Methods:The CYP1A1 gene polymorphism(an T-C transversion at nucleotide position 3801)was detected by the polymerase chain reaction and restriction fragment length polymorphism in 80 cases with breast cancer and 60 samples of normal breast tissue.The difference in genotypic distribution frequency between the groups,the correlation between the genotypes and the factors related to prognosis were analyzed.Results:The incidence of homozygous and variant genotypes had no difference between the breast cancer group and controls group(P=0.746).The proportion of variant genotype increased as clinical stage(P=0.006)advanced,as well as with increased numbers of lymph node metastases(P=0.010).Conclusions:In patients with breast cancer there is a correlation between the CYP1A1 CC allele and some factors indicating poor prognosis,including more lymph node metastases as well as a more advanced clinical stage.

Mitochondrial malic enzyme 2 promotes breast cancer metastasis via stabilizing HIF-1α under hypoxia

Objective: α-ketoglutarate(α-KG) is the substrate to hydroxylate collagen and hypoxia-inducible factor-1α(HIF-1α), which are important for cancer metastasis. Previous studies have shown that the upregulation of collagen prolyl 4-hydroxylase in breast cancer cells stabilizes the expression of HIF-1α by depleting α-KG levels. We hypothesized that mitochondrial malic enzyme 2(ME2) might also affect HIF-1α expression via modulating α-KG levels in breast cancer cells.Methods: We evaluated ME2 protein expression in 100 breast cancer patients using immunohistochemistry and correlated with clinicopathological indicators. The effect of ME2 knockout on cancer metastasis was evaluated using an orthotopic breast cancer model. The effect of ME2 knockout or knockdown on the levels of α-KG and HIF-1α proteins in breast cancer cell lines was determined both in vitro and in vivo.Results: ME2 was found to be upregulated in the human breast cancerous tissues compared with the matched precancerous tissues(P<0.001). The elevated expression of ME2 was associated with a poor prognosis(P=0.019).ME2 upregulation was also related to lymph node metastasis(P=0.016), pathological staging(P=0.033), and vascular cancer embolus(P=0.014). Also, ME2 knockout significantly inhibited lung metastasis in vivo. In the tumors formed by ME2 knockout cells, the levels of α-KG were significantly increased and collagen hydroxylation level did not change significantly but HIF-1α protein expression was significantly decreased, compared to the control samples. In cell culture, cells with ME2 knockout or knockdown demonstrated significantly higher α-KG levels but significantly lower HIF-1α protein expression than control cells under hypoxia. Exogenous malate and α-KG exerted similar effect on HIF-1α in breast cancer cells to ME2 knockout or knockdown. Additionally,treatment with malate significantly decreased 4 T1 breast cancer lung metastasis. ME2 expression was associated with HIF-1α levels in human breast cancer samples(P=0.008).Conclusions: Our results provide evidence that upregulation of ME2 is associated with a poor prognosis of breast cancer patients and propose a mechanistic understanding of a link between ME2 and breast cancer metastasis.

miR-200c Inhibits Metastasis of Breast Cancer Cells by Targeting HMGB1

miR-200c has been shown to regulate the epithelial-mesenchymal transition （EMT） by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software （miRanda） was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB 1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of＇miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

Nef-M1, a CXCR4 Peptide Antagonist, Enhances Apoptosis and Inhibits Primary Tumor Growth and Metastasis in Breast Cancer

Results from studies with animal models suggest that, in many cancers, CXCR4 is an important therapeutic target and that CXCR4 antagonists may be promising treatments for primary cancers and for metastases. The Nef protein effectively competes with CXCR4’s natural ligand, SDF-1α, and induces apoptosis. As described in this report, the Nef-M1 peptide (Nef protein amino acids 50 - 60) inhibits primary tumor growth and metastasis of breast cancer (BC). Four BC cell lines (MDA-MB-231, MDA-MB-468, MCF 7, and DU4475) and primary human mammary epithelium (HME) cells were evaluated for their response to the Nef protein and to the Nef-M1 peptide. The presence of CXCR4 receptors in these cells was determined by RT-PCR, Western blot (WB), and immunohistochemical analyses. The apoptotic effect of Nef-M1 was assessed by terminal transferase dUTP nick-end labeling (TUNEL). WBs was used to assess caspase 3 activation. BC xenografts grown in SCID mice were evaluated for the presence of CXCR4 and for their metastatic potential. CXCR4 was presented in MDA-MB-231, MCF 7, and DU 4475 BC cells but not in MDA-MB-468 BC or HME cells. Cells expressing CXCR4 and treated with Nef-M1 peptide or the Nef protein had higher rates of apoptosis than untreated cells. Caspase-3 activation increased in MDA-MB 231 cells treated with the Nef protein, the Nef 41 - 60 peptide, or Nef-M1. Nef-M1, administered to mice starting at the time of xenograft implantation, inhibited growth of primary tumors and metastatic spread. Untreated mice developed diffuse intraperitoneal metastases. We conclude that, in BCs, Nef-M1, through interaction with CXCR4, inhibits primary tumor growth and metastasis by causing apoptosis.

Serum Y-Box Binding Protein 1 (YBX-1) and Interleukin 6 (IL-6) Are Associated with Metastasis in Breast Cancer Patients

Objectives: The aim of this study was to assess the levels of Y-box binding protein 1 (YBX-1) and interleukin 6 (IL-6) in the sera of metastatic and non-metastatic breast cancer patients (BC), investigate their clinicopathological significance and to analyze their potential use as biomarkers of breast cancer metastasis. Methods: The study included ninety subjects sub-grouped equally into metastatic BC, non-metastatic BC and healthy volunteers. Serum YBX-1 and IL-6 were quantified using ELISA technique while CA 15-3 was quantified using IRMA kit. Clinical data were collected from patients’ records. Results: YBX-1 (p < 0.001), IL-6 (p < 0.001) and CA15-3 (p = 0.017, 0.001) were significantly elevated in metastatic and non-metastatic BC patients compared to healthy controls, however, only YBX-1 (p 0.001) showed a significant difference with cancer metastasis. Generally, YBX-1 and IL-6 were correlated with worse histological grade and late clinical stage in breast cancer patients and they were also associated with axillary lymph nodes involvement and positive vascular invasion in metastatic BC patients. Serum YBX-1 and IL-6 levels were positively correlated to each other (rs = 0.615, p < 0.001) and they showed high sensitivity and specificity compared to CA 15-3 (p < 0.001 and p = 0.004 for YBX-1 and IL-6 respectively) for predicting cancer metastasis. Conclusions: Serum YBX-1 and IL-6 are potential biomarkers of breast cancer patients with significant correlation with bad clinicopathological characteristics. Serum YBX-1 and IL-6 have superior sensitivity and specificity compared to CA15-3 and can serve as potential follow up and prognostic markers.

DOT1L-long enhances breast cancer metastasis

Objective· To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis. Methods· The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition(EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high. Results· PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells. Conclusion· The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.

Breast metastasis from lung cancer：a report of two cases and literature review

Breast metastasis from extra-mammary malignancy is rare. An incidence of 0.4% to 1.3% has been reported in literature. The primary malignancies that most commonly metastasize to the breast are leukemia, lymphoma, and malignant melanoma. In this report, two cases of pulmonary metastasis to the breast were presented. A 40-year-old female manifested a right breast mass of 2-month duration. After physical examination was performed, a poorly defined mass was noted in the upper outer quadrant of the right breast. Another 49-year-old female manifested right breast mass of 5-day duration. A poorly defined mass was noted in the lower inner quadrant of the right breast. Mammography results also revealed breast cancer. The patients underwent local excision. After histological and immunohistochemical analyses were conducted, a primary lung carcinoma that metastasized to the breast was diagnosed. An accurate differentiation of metastasis to the breast from primary breast cancer is very important because the treatment and prognosis of the two differ significantly.

G Protein-Coupled Estrogen Receptor Is a Critical Regulator in Metastasis of Breast Cancer Cells

Estrogen signaling via GPER in breast cancers has been intensively discussed over years, but the underlying molecular mechanism remains to be fully elucidated, especially for the transcriptional profiles of the GPER under estrogen stimulation. In this study, we evaluated the potential role GPER in regulating invasion of metastatic breast cancer cell line MDA-MB-231 via transcriptional way. First, with primary breast cancer tissue samples, we found that the expression of GPER significantly coordinated with another membrane receptor-CXCR1. Besides, the expression level of these two proteins was associated with the development of the primary breast cancers. Second, to dissect the cross talk between GPER and CXCR1, we further our studies to detect the activation of ERK, Akt and transcriptional factor NF-kB. We found that upon estrogen stimulation, the phosphorylation level of ERK and Akt were rapidly increased and then resulted in the activation and nucleus translocation of NF-kB. When we considered the sub-sequence of the NF-kB activation, we found the autocrine of IL-8 was boosted by stimulation of the estrogen and followed by the promoted invasion of the MDA-MB-231 cells. In conclusion, our data demonstrated the estrogen-mediated GPER stimulation would regulate the invasive activities of metastatic breast cancer cell line MDA-MB-231 coupled with another membrane receptor CXCR1 via transcriptional pathway, the cross talk between GPER and CXCR1 may be another target for breast cancer therapies.

血清外泌体SUMO1P3和MALAT1对三阴性乳腺癌患者术后复发转移的预测价值

目的探讨血清外泌体小泛素样修饰子1伪基因3(SUMO1P3)和肺腺癌转移相关转录本1(MALAT1)对三阴性乳腺癌(TNBC)患者术后复发转移的预测价值。方法选取2016年至2020年行手术切除的224例TNBC患者作为研究对象,根据随访期内复发及转移情况分为非复发转移组和复发转移组,收集两组患者的临床病历资料。采用实时荧光定量PCR(qPCR)检测术前血清外泌体SUMO1P3和MALAT1表达,绘制受试者工作特征(ROC)曲线评价SUMO1P3和MALAT1对TNBC术后复发转移的预测价值。采用多因素Logistic回归模型分析影响TNBC术后复发转移的因素。结果共72例患者出现复发或转移。224例TNBC患者SUMO1P3和MALAT1相对表达量分别为3.06±0.68和2.75±0.66,其中复发转移组SUMO1P3和MALAT1相对表达量为3.44±0.67和3.46±0.80,均显著高于非复发转移组的2.88±0.61和2.33±0.48(P<0.001)。ROC曲线结果显示,血清外泌体SUMO1P3和MALAT1联合检测的曲线下面积(AUC)=0.842(95%CI:0.787~0.887),特异度为0.790,灵敏度为0.806,且两指标联合预测的效能优于SUMO1P3和MALAT1单独预测(P<0.05)。多因素Logistic回归模型结果显示,SUMO1P3(OR=4.463,95%CI:2.125~9.372)和MALAT1(OR=9.103,95%CI:4.462~18.573)表达是影响TNBC术后复发转移的独立因素(P<0.05)。结论血清外泌体SUMO1P3和MALAT1与TNBC患者预后密切相关,两指标联合检测对TNBC术后复发或转移具有较高的预测价值。

Effects of Notch-1 Down-regulation on Malignant Behaviors of Breast Cancer Stem Cells

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells （BCSCs）. BCSCs were enriched by using serum-free me- dium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and fl0w cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a prom- ising therapeutical approach for breast cancer.

MicroRNA-6744-5p promotes anoikis in breast cancer and directly targets NAT1 enzyme

Objective:Anoikis is apoptosis that is induced when cells detach from the extracellular matrix and neighboring cells.As anoikis serves as a regulatory barrier,cancer cells often acquire resistance towards anoikis during tumorigenesis to become metastatic.MicroRNAs(miRNAs)are short strand RNA molecules that regulate genes post-transcriptionally by binding to mRNAs and reducing the expression of its target genes.This study aimed to elucidate the role of a novel miRNA,miR-6744-5 p,in regulating anoikis in breast cancer and identify its target gene.Methods:An anoikis resistant variant of the luminal A type breast cancer MCF-7 cell line(MCF-7-AR)was generated by selecting and amplifying surviving cells after repeated exposure to growth in suspension.Mi RNA microarray analysis identified a list of dysregulated miRNAs from which miR-6744-5 p was chosen for overexpression and knockdown studies in MCF-7.Additionally,the miRNA was also overexpressed in a triple-negative breast cancer cell line,MDA-MB-231,to evaluate its ability to impair the metastatic potential of breast cancer cells.Results:This study showed that overexpression and knockdown of miR-6744-5 p in MCF-7 increased and decreased anoikis sensitivity,respectively.Similarly,overexpression of miR-6744-5 p in MDA-MB-231 increased anoikis and also decreased tumor cell invasion in vitro and in vivo.Furthermore,NAT1 enzyme was identified and validated as the direct target of miR-6744-5 p.Conclusions:This study has proven the ability of miR-6744-5 p to increase anoikis sensitivity in both luminal A and triple negative breast cancer cell lines,highlighting its therapeutic potential in treating breast cancer.

剪接因子3B亚单位1在乳腺癌组织中的表达及与患者临床病理特征关系

目的:探讨剪接因子3B亚单位1(SF3B1)在乳腺癌组织中的表达及与患者临床病理特征的关系。方法:分析88例乳腺癌患者乳腺癌组织和癌旁组织SF3B1的表达水平,评估SF3B1对乳腺癌的诊断价值,比较SF3B1高表达组与低表达组临床病理特征差异。结果:乳腺癌组织SF3B1表达明显高于癌旁组织(P<0.05);SF3B1诊断乳腺癌的AUC为0.713;SF3B1诊断乳腺癌的最佳截断值为110.72。高表达组年龄明显高于低表达组,淋巴结转移为N 0的比例明显低于低表达组,两组病理学分级比较差异有统计学意义(P<0.05)。结论:SF3B1在乳腺癌组织中的表达明显上调,在诊断乳腺癌方面效能较好,与临床病理参数有关,可能参与了乳腺癌的发生、侵袭和转移过程的调控。

食管鳞癌组织中BARD1的表达及与淋巴结转移的关系

目的分析食管鳞癌(ESCC)中乳腺癌易感基因1相关环指结构域蛋白1(BARD1)的表达及与淋巴结转移的关系。方法纳入2018年1月至2021年12月本院收治且经组织病理学检查确诊的114例ESCC患者,免疫组化EnVision两步法检测癌组织与癌旁组织中BARD1的表达情况并比较BARD1阳性率,分析BARD1表达与ESCC临床病理特征的关系,建立Logistic回归模型分析BARD1表达对ESCC淋巴结转移的影响,绘制受试者工作特征(ROC)曲线分析BARD1表达对ES⁃CC淋巴结转移的预测价值。结果114例ESCC组织中BARD1阳性78例、阴性36例,对应癌旁组织中BARD1阳性23例、阴性91例,ESCC组织的BARD1阳性率为68.42%,高于癌旁组织的20.18%(χ^(2)=53.769,P<0.001)。BARD1表达与ESCC患者的T分期、分化程度和淋巴结转移有关(P<0.05),其中淋巴结转移组织的BARD1阳性率为80.36%(45/56),高于未转移组织的56.90%(33/58)。经Phi系数分析发现,BARD1表达与ESCC淋巴结转移呈正相关(r=0.276,P=0.003)。BARD1阳性是ESCC患者淋巴结转移的独立危险因素(OR=3.099,95%CI:1.339~7.175,P=0.008),且BARD1表达对ESCC患者淋巴结转移具有一定预测价值(AUC=0.636,P=0.020)。结论BARD1在ESCC中高表达,其与ESCC淋巴结转移密切相关,BARD1阳性会增加淋巴结转移风险,在评估ESCC淋巴结转移上有一定价值。

Prognostic significance of MDR-1 P-glycoprotein expression in breast cancer

Objective： To investigate the expression of MDR-1 P-glycoprotein（MDR-1 Pgp） in breast cancer and analyze its correlation to the biological behavior and prognosis of the disease. Methods：The expression of MDR-1 Pgp was examined in 75 cases of breast cancer patients by using three different monoclonal antibodies（JSB1, C219 and C494） with S-P immunohistochemisty. These patients were followed up for 5 years, and the correlation between MDR-1 Pgp expression, survival rate and lymph metastasis was analyzed. Results： Positive detection of MDR-1 Pgp by JSB1, C219 and C494 in 75 cases of breast cancer was 86.7%, 48% and 85.3%, respectively. MDR-1 Pgp expression was not related to ages of patients （P 〉 0.05）. JSBl-detected expression of MDR-1 Pgp was related to lymph node metastasis（P〈 0.05）; while C219 and C494 were not（P 〉 0.05）. The patients with MDR-1 Pgp expression positively detected by either two of the three antibodies, had five-year survival rate that was significantly higher than those positively detected by all the three antibodies（P 〈 0.05）. Conclusion：Three antibodies should be used simultaneously to detect MDR-1 Pgp expression in breast cancer. Positive MDR-1 Pgp expression in breast cancer detected by all the three antibodies may represent a poor prognosis; while positive MDR-1 Pgp detection by JSB1 and C494 is associated with lymph metastasis.

Evaluation of imatinib mesylate(Gleevec) on KAI1/CD82 gene expression in breast cancer MCF-7 cells using quantitative real-time PCR

Objective: To evaluate the effect of imatinib mesylate on cell viability, anti cancer effect through modulation of KAI1/CD82 gene expression in breast cancer MCF-7 cell line.Methods: The effects of imatinib mesylate on cell viability in MCF-7 cell line were assessed using MTT assay and IC_(50) value was determined. GAPDH and KAI1/CD82 were selected as reference and target genes, respectively. Quantitative real time PCR technique was applied for investigation of KAI1/CD82 gene expression in human breast cancer MCF-7 cells. Subsequently, the quantity of KAI1 compared to GAPDH gene expressions were analyzed using the formula; 2^(-DDCt).Results: Imatinib was showed to have a dose-dependent inhibitory effect on the viability of MCF-7 cells. CD82/GAPDH gene expression ratios were 1.322 ± 0.030(P > 0.05),2.052 ± 0.200(P < 0.05), 2.151 ± 0.270(P < 0.05) for 10, 20 and 40 mmol/L of imatinib concentrations.Conclusions: Based on the present data, imatinib mesylate might modulate metastasis by up-regulating KAI1/CD82 gene expression in human breast MCF-7 cancer cell line.

MicroRNA-26b下调MALAT-1抑制乳腺癌MCF-7细胞恶性生物学行为的机制研究

目的 探索microRNA-26b(miR-26b)通过MALAT-1抑制乳腺癌MCF-7细胞恶性生物学行为的分子机制。方法 以乳腺癌细胞系MCF-7作为研究对象,采用慢病毒LV-miR-26b-ctrl和LV-miR-26b转染肿瘤细胞,逆转录聚合酶链反应检测MALAT-1、miR-26a和miR-26b的mRNA表达,平板克隆形成实验、CCK-8细胞增殖实验、软琼脂成瘤实验、细胞划痕实验、Transwell细胞侵袭实验分别检测肿瘤细胞的增殖、体外成瘤、迁移和侵袭能力。结果 E2组与Mock组MCF-7细胞24、48、72和96 h时吸光度值比较,经重复测量设计的方差分析,结果:(1)不同时间点吸光度值比较,差异有统计学意义(P <0.05);(2)两组吸光度值比较,差异有统计学意义(P <0.05),与Mock组比较,E2组72和96 h时细胞增殖能力较Mock组提高(P <0.05);(3)两组吸光度值变化趋势比较,差异有统计学意义(P <0.05)。与Mock组相比,E2组MALAT-1相对表达量升高(P <0.05),miR-26a、miR-26b mRNA相对表达量下降(P <0.05)。高表达miR-26b的乳腺癌患者的生存情况优于低表达miR-26b组,死亡风险更低(P <0.05),低表达miR-26a组患者的生存情况优于高表达miR-26a组(P <0.05),用Cox比例风险回归模型,将miR-26a作为一个独立的预后因素来比较,两组患者的死亡风险比较无差异(P>0.05)。与LV-miR-26b-ctrl组比较,LV-miR-26b-ctrl/E2组乳腺癌细胞的MALAT-1相对表达量升高(P <0.05),与LV-miR-26b-ctrl/E2组比较,LV-miR-26b/E2组乳腺癌细胞的MALAT-1相对表达量降低(P <0.05)。LV-miR-26b-ctrl组、LV-miR-26b-ctrl/E2组、LV-miR-26b/E2组细胞在24、48、72和96 h时吸光度值比较,经重复测量设计的方差分析,结果:(1)不同时间点吸光度值比较,差异有统计学意义(P <0.05);(2)各组吸光度值比较,差异有统计学意义(P <0.05),LV-miR-26b-ctrl/E2组72和96 h时细胞增殖能力明显较LV-miR-26b-ctrl组增强(P <0.05),LV-miR-26b/E2组72和96 h时细胞增殖能力较LV-miR-26b-ctrl/E2组降低(P <0.05);(3)各组吸光度值变化趋势比较,差异有统计学意义(P <0.05)。E2处理后的LV-miR-26b-ctrl肿瘤细胞增殖和体外成瘤能力增强。与LV-miR-26bctrl/E2组比较,LV-miR-26b/E2组肿瘤细胞的增殖和体外成瘤能力降低。在24 h时,LV-miR-26b-ctrl/E2组细胞划痕间隙较LV-miR-26b-ctrl组变窄,LV-miR-26b/E2组24 h时细胞的划痕间隙较LV-miR-26bctrl/E2组明显增宽。LV-miR-26b-ctrl/E2组Transwell小室下方的乳腺癌细胞数量较LV-miR-26b-ctrl组增多,LV-miR-26b/E2组的肿瘤细胞数量较LV-miR-26b-ctrl/E2组减少。结论 下调MALAT-1可能是miR-26b抑制乳腺癌恶性生物学行为的分子机制之一,miR-26b有望成为乳腺癌治疗的调控靶点。

血清LRG1表达水平在乳腺癌诊断中的应用

目的:检测乳腺癌患者血清和癌组织内富亮氨酸α2糖蛋白1(leucine-richalpha-2glycoprotein1,LRG1)的表达并分析其临床意义。方法:选取2021年6月—2022年3月就诊于牡丹江市肿瘤医院的86例女性乳腺癌患者,86例乳腺小叶增生或纤维瘤患者作为良性病变组,21例健康志愿者作为对照组。比较三组血清LRG1的表达差异,分析其与患者年龄、临床分期及淋巴结转移等临床因素的关系。免疫组化染色检测癌组织和癌旁组织内LRG1表达的平均灰度值。结果:乳腺癌组和良性病变组血清内LRG1的表达均明显高于对照组,乳腺癌组LRG1表达明显高于良性病变组,差异均有统计学意义(P<0.05)。不同年龄组乳腺癌患者血清内LRG1的表达,差异有统计学意义(P<0.05)。Ⅰ+Ⅱ期乳腺癌患者血清内LRG1的表达低于Ⅲ+IV期,差异有统计学意义(P<0.05)。淋巴结转移组血清内LRG1表达明显高于未发生淋巴结转移组,差异有统计学意义(P<0.05)。乳腺癌组织内LRG1平均灰度值明显低于癌旁组织,差异有统计学意义(P<0.05)。结论:LRG1在乳腺癌患者血清和癌组织内明显升高,并与临床因素紧密相关,可作为乳腺癌诊断的潜在生物标志物。

超声测定PI、RI、Vmax结合血清sE-cadherin、CYFRA21-1对乳腺癌良恶性及淋巴结转移的诊断价值

目的研究超声测定的搏动指数(PI)、血流阻力指数(RI)、最大血流值(Vmax)结合血清可溶性E-钙黏连蛋白(sE-cadherin)、细胞角蛋白19片段(CYFRA21-1)对乳腺癌良恶性及腋窝淋巴结转移的诊断价值。方法选择242例乳腺肿块女性患者,年龄43~67岁,平均年龄55.23岁;身体质量指数19.5~28.9 kg/m^(2),平均身体质量指数24.12 kg/m^(2);病程4~8个月,平均病程6.45个月;均为初次入院就诊。彩色多普勒超声仪测定PI、RI和Vmax,并在术前采集空腹外周血,酶联免疫吸附分析测定血清sE-cadherin和CYFRA21-1。根据病理组织诊断结果将患者分成乳腺癌组(n=132)和乳腺良性组(n=110),并将乳腺癌患者分成腋窝淋巴结转移组(n=74)和腋窝淋巴结未转移组(n=58)。单因素分析临床资料,Logistic回归分析乳腺癌发生和腋窝淋巴结转移的独立影响因素,通过受试者工作特性(ROC)曲线分析各指标及回归分析诊断效能。结果乳腺良恶性病变区分ROC曲线结果表明,PI临界值1.34,灵敏度为69.70%,特异度为70.91%,曲线下面积(AUC)为0.801[95%可信区间(CI)0.750~0.852];RI诊断的临界值为0.69,灵敏度为65.15%,特异度65.45%,AUC为0.762(95%CI 0.707~0.812);Vmax诊断的临界值为13.11 m/s,灵敏度为59.85%,特异度60.00%,AUC为0.627(95%CI 0.564~0.699);血清sE-cadherin诊断的临界值为24.18 ng/mL,灵敏度为62.88%,特异度58.18%,AUC为0.709(95%CI 0.645~0.771);血清CYFRA21-1诊断的临界值为9.55 g/mL,灵敏度为74.24%,特异度75.45%,AUC为0.817(95%CI 0.769~0.865)。将回归预测方程作为新变量P,在最佳临界切点时,回归分析灵敏度81.82%,特异度79.09%,AUC为0.867(95%CI 0.829~0.908),回归分析灵敏度显著高于各指标单独检测(P<0.05)。乳腺癌淋巴结转移分析ROC曲线结果表明,PI临界值1.58,灵敏度为79.73%,特异度为68.97%,AUC为0.815(95%CI 0.727~0.902);RI诊断的临界值为0.83,灵敏度为63.51%,特异度70.69%,AUC为0.701(95%CI 0.558~0.844);Vmax诊断的临界值为17.36 m/s,灵敏度为54.05%,特异度55.17%,AUC为0.516(95%CI 0.384~0.649);血清sE-cadherin诊断的临界值为34.52 ng/mL,灵敏度为82.43%,特异度60.34%,AUC为0.755(95%CI 0.653~0.856);血清CYFRA21-1诊断的临界值为13.10 g/mL,灵敏度为60.81%,特异度65.52%,AUC为0.563(95%CI 0.426~0.701)。将回归预测方程作为新变量P,在最佳临界切点时,回归分析灵敏度85.14%,特异度84.48%,AUC为0.893(95%CI 0.816~0.970),回归分析灵敏度显著高于各指标单独检测(P<0.05)。结论超声参数及血清sE-cadherin和CYFRA21-1在乳腺癌和腋窝淋巴结中异常表达,且PI、RI、Vmax和血清sE-cadherin、CYFRA21-1单独和Logistic回归分析可应用于乳腺恶性病变和淋巴结转移的预测和诊断。

乳腺癌患者肿瘤转移相关基因1表达与化疗敏感性及预后的相关性

目的研究乳腺癌患者肿瘤转移相关基因1(MTA1)表达与化疗敏感性及预后的相关性。方法选取92例接受乳腺癌根治术的患者作为研究对象,根据患者术后化疗结果分为敏感组和抵抗组,比较2组患者的MTA1表达水平以及相关病理参数,采用logistic回归分析影响患者化疗敏感性的因素,同时根据随访结果分析MTA1表达水平与患者预后的相关性。结果92例化疗患者中敏感组患者共53例,抵抗组共39例。敏感组患者MTA1高表达率低于抵抗组(P<0.05)。敏感组患者中年龄<55岁、KPS评分≥80分、肿瘤分期Ⅰ/Ⅱ期患者占比高于抵抗组(P<0.05)。Logistic回归分析结果显示年龄≥55岁、Karnofsky功能状态评分(KPS)<80分、肿瘤分期Ⅲ期以及MTA1高表达状态均是影响化疗敏感性的高危因素(P<0.05)。MTA1高表达组中位PFS为8个月以及中位OS为13个月,低表达组的中位PFS为12个月以及中位OS为16.5个月,差异比较具有统计学意义(P<0.05)。结论乳腺癌患者MTA1表达与化疗效果有关,其高表达提示化疗不敏感和预后不良,可能成为预测患者预后的重要指标。

In Vivo Animal Model Evaluation of a Powerful Oral Nanomedicine for Treating Breast Cancer in BALB/c Mice Using 4T1 Cell Lines without Chemotherapy

Nanopharmaceuticals containing quantum dot nanoparticles (Q-Dot NPs) for treating serious cancers such as breast cancer have made fantastic proposals. In this study, ZnO quantum dot NPs are formulated via ZnO@PVP nanopolymer as co-assistants coordinating with efficacious suitable wetting agents, PEG-binding compound, and W/O emulsifier for producing eco-friendly water-based nanodrug. Several characterization techniques containing SEM, TEM, FTIR, photoluminescence, zeta potential, and UV-Vis absorption were employed for ZnO Q-Dot NPs in nanodrug. This work aims to investigate the anti-tumor effects of such nanomedicine on the 4T1 breast cancer cell line in BALB/c mice, being elaborated through intraperitoneal, injection (IVP) and oral therapy. The impressive findings showed that ZnO nanodrug caused changes in blood factors, having the most effectiveness at 40 μg/ml concentration after two weeks of oral treatments. The significant increase in white blood cells (WBC) neutrophils and meaningful decreases in lymphocytes and especially cholesterol were powerful simultaneous impacts, successfully treating malignant breast cancer masses. In this significant animal model research for breast cancer, the sick mice recovered entirely and even had a safe space to mate. Histopathological results showed no evidence of breast tumor formation or metastasis in the group treated with nanodrug and their children. This nanomedicine has a therapeutic effect, and is ready to be applied for treating volunteer breast cancer patients. However, its prevention (inhibitory) effect can also be analyzed and added to current data in future studies.