PurposeRecommendations from the National Health Commission of China (NHCC) and the International Ki67 Working Group (IKWG) were issued to guide immunohistochemistry (IHC)-based Ki67 scoring for breast cancer patients in daily clinical practice. They were evaluated in this multi-institutional study alongside the results from the Quantitative Dot Blot (QDB) method.MethodsThree alternative adjacent sections from 40 primary ER+ breast cancer resection blocks were randomly assigned a number from 1 to 120 for Ki67 staining and reviewed by 21 pathologists, while the other three alternative sections were sent for QDB analysis of Ki67 protein levels. Ki67 scores were grouped by 5/30% (IKWG), 10/30% (NHCC) and 20/30% (NHCC appendix 9, NHCCa9), respectively while QDB results were grouped by C5–C95 of 2.31 nmol/g defined in previous study as low-, equivocal-, and high-risk groups.ResultsThe overall Intraclass Correlation Coefficient (ICC) was 0.785 for IHC evaluations from 21 pathologists, with Fleiss Kappa values of 0.555, 0.628, and 0.480 when Ki67 scores were grouped by guidance from IKWG, NHCC, and NHCCa9, respectively. In comparison, the ICC and Fleiss kappa values for the QDB analysis were 0.939 and 0.831, respectively. When IHC and QDB results were cross-referenced, more specimens were grouped as high-risk by QDB than IHC, and NHCCa9 led to the highest percentage of disagreement between the two methods.ConclusionThe IKWG recommendation was harder to achieve categorized agreement among pathologists than the NHCC recommendation, yet it led to the best agreement with the QDB to define the low-risk group. The QDB method offers significantly improved consistency compared to the current IHC-based Ki67 assessment.

Background: Choriocarcinoma (CC), a highly aggressive and malignant subtype of gestational trophoblastic disease (GTD), arises from the dysregulated proliferation of trophoblastic cells, which normally mediate placental development during pregnancy. This study aimed to characterize the expression and functional significance of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), a nuclear-enriched long non-coding RNA, in CC pathogenesis. Methods: We analyzed MALAT1 expression levels in 30 normal placental villi, 46 hydatidiform moles (repressive subtype), and 52 CC specimens. To identify MALAT1-interacting proteins, we performed RNA antisense purification followed by immunoblotting, with subsequent validation via co-immunoprecipitation (Co-IP) and immunofluorescence (IF) assays. Results: MALAT1 was significantly upregulated in CC tissues compared to controls. In vivo xenograft experiments further revealed that MALAT1 overexpression enhanced tumor growth. Mechanistically, we identified RNA-binding motif protein 10 (RBM10), a key spliceosomal regulator, as a novel MALAT1-binding partner. Strikingly, MALAT1 promoted p53 protein degradation without affecting its transcriptional levels, and this oncogenic effect was mediated through an RBM10-dependent mechanism. Conclusions: In conclusion, our findings establish MALAT1 as a critical oncogenic driver in CC, functioning through its interaction with RBM10 to destabilize p53 and accelerate tumor progression. These insights highlight the potential of the MALAT1-RBM10-p53 axis as a therapeutic target in CC.

Abstract Purpose: Recommendations from National Health Commission of China (NHCC) and International Ki67 Working Group (IKWG) were issued respectively to guide immunohistochemistry (IHC)-based Ki67 scoring for breast cancer patients in daily clinical practice. They were evaluated in this multi-institutional study alongside with results from Quantitative Dot Blot (QDB) method. Method: Three sections each from 40 primary ER+ breast cancer resection blocks were randomly assigned a number from 1 to 120 for Ki67 staining and reviewed by 21 pathologists while the other three alternative sections were sent for QDB analysis of Ki67 protein levels. Ki67 scores were grouped by 5/30% (IKWG), 10/30% (NHCC) and 20/30% (NHCC appendix 9, NHCCa9) respectively while QDB results were grouped by C5-C95 of 2.31 nmole/g defined in previous study as low, intermediate and high risk groups. Results: The overall Intraclass Correlation Coefficient (ICC) was 0.785 for IHC evaluations from 21 pathologists, with the Fleiss Kappa at 0.555, 0.628 and 0.480 when Ki67 scores were grouped by the guidance from IKWG, NHCC and NHCCa9 respectively. In comparison, the ICC and Fleiss kappa for QDB analysis were at 0.939 and 0.831. When IHC and QDB results were cross-referenced, more specimens were grouped as high risk by QDB than IHC, and NHCCa9 led to highest percentage of disagreement between two methods. Conclusion: The IKWG recommendation was harder to achieve categorized agreement among pathologists than that of NHCC, yet it led to best agreement with QDB to define low-risk group. QDB method offered significantly improved consistency over current IHC-based Ki67 assessment.

BACKGROUND:To establish the pathological diagnosis of UTUC before treatment is profitable. At present, the conventional pathological diagnostic methods have certain problems. Besides, the urine-based DNA methylation test have been already utilized to detect bladder cancer. OBJECTIVE:To evaluate the sensitivity and specificity of DNA methylation plus 17 genes mutation test and compare the combined test with cytology. MATERIALS AND METHODS:We included 45 patients from April 2019 to May 2022, all of whom underwent radical nephroureterectomy (RNU), nephrectomy, diagnostic ureteroscopy or tissue biopsy. Before surgery, the urine samples were collected for DNA methylation plus 17 genes mutation test and cytology. The test performance was calculated, and comparative ROC curves were drawn. RESULTS:The median age of the patients was 67 years. The Kappa value of the DNA methylation plus 17 genes mutation test and tissue pathology was 0.59 (p<0.001). The sensitivity/specificity/PPV/NPV of DNA methylation plus 17 genes mutation test was 86/80/94/62% compared with 29/100/100/29% for cytology. The AUC of DNA methylation plus 17 genes mutation test was 0.829 (p<0.001).The mutated gene proportion of UTUC patients was 51.43% for TERT and 25.71% for TP53. CONCLUSION:The test performance of DNA methylation plus 17 genes mutation test was satisfactory, which may replace cytology in the future. Further multicenter studies with larger samples are needed to confirm the clinical value of this promising method. NOVELTY & IMPACT STATEMENTS:We evaluated the diagnostic efficacy of a urine-based liquid biopsy for the detection of UTUC and compared the combined test with cytology. We found satisfactory results and concluded that the test could partly replace cytology. Further studies are needed.

This study aimed to analyze the relationship between human epididymal protein 4 (HE4) and infiltration depth, postoperative recurrence, and metastasis of epithelial ovarian cancer (OVCA). Immunohistochemistry was used to detect the expression level of HE4 in cancer tissues and adjacent tissues of 90 patients with epithelial OVCA admitted to our hospital from May 2017 to January 2018. Cox regression was used to analyze the factors affecting the prognosis of epithelial OVCA. The relationship between HE4 and the prognosis of epithelial OVCA was analyzed by the receiver operating characteristic curve and Kaplan-Meier survival curve. The positive expression rate of HE4 in epithelial OVCA was 85.56%, which was higher than 34.44% in adjacent tissues (p < 0.01). The International Federation of Gynecology and Obstetrics stage, infiltration depth, lymph node metastasis, postoperative recurrence and metastasis, and HE4 positivity were independent risk factors for the prognosis, and platinum-based chemotherapy sensitivity was an independent protective factor for the prognosis of patients with epithelial OVCA (p < 0.05). The area under the curve of HE4 in diagnosing epithelial OVCA and predicting recurrence was 0.863 and 0.700, the sensitivity was 91.60% and 85.60%, and the specificity was 90.20% and 65.60%. The median progression-free survival and overall survival were 26.1 and 30.2 months in HE4-positive epithelial OVCA patients, while these were 31.4 and 35.6 months in HE4-negative epithelial OVCA patients (p < 0.05). In conclusion, HE4 was highly expressed in epithelial OVCA tissues. Its expression level was related to the depth of tumor invasion, postoperative recurrence and metastasis, and other clinicopathological characteristics of patients with epithelial OVCA.

腺泡状软组织肉瘤( alveolar soft-part sarcoma, ASPS)是一种组织来源至今未明确的恶性软组织肿瘤,有文献[1]报道考虑颗粒细胞肌母细胞来源、血管平滑肌细胞来源、肾小球旁细胞来源.ASPS好发于15～35岁女性,在儿童中多见于头部和颈部,成年人常见于下肢深部软组织,好发于右侧肢体[2].国内外也有少数报道[3-8]发生于颅内、舌、口咽、头颈、乳腺、子宫体、宫颈等部位,目前仅有英文文献[9-12]报道8 例原发于阴道,国内文献未检索到发生于阴道壁的病例.现报道河南省人民医院收治的阴道壁ASPS 1例,并结合国内外相关文献,分析该病的诊治特点,以提高妇科医师对该病的认识.

Objective:To investigate the MRI features of fibro-adipose vascular anomaly (FAVA).Methods:The clinical, pathological and MRI data of the FAVA patients confirmed clinically and pathologically in Henan Provincial People’s Hospital from January 2016 to June 2021 were retrospectively analyzed, including 17 females and 13 males, aged 4-53 (18.5±12.8) years.Through the analysis of the MRI images of FAVA performance, including the lesion area, shape, signal, the degree of reinforcement, inner structure, outer structure and other image characteristics, summarize the MRI features.Results:The lesions located at the lower leg (12 cases), thigh (12 cases), upper arm (4 cases), forearm (1 case) and trunk (1 case). Twenty-four case of lower limbs. All cases involved the muscular layer, including 21 cases of superficial muscle layer, 7 cases of deep muscle layer, and 2 cases of both. Twenty-three cases of superficial muscle layer. Most of the affected muscles were quadriceps femoris and gastrocnemius. Morphology: all lesions were intramuscular solid masses growing parallel to the long axis of the muscle. There were 11 cases of focal type, 11 cases of locally infiltrating type, and 8 cases of diffuse type. MRI findings: all lesions showed mixed signal. On T1WI, 28 cases showed cloud-like, band-like and patchy high signal on the background of medium and high signal. On T2WI-FS, all the 30 cases showed low to medium signal areas on the background of high signal, which were dendrimer, ribbon and cloud-like. The lesions showed moderate to obvious heterogeneous progressive enhancement. Twenty-seven lesions had different shapes of vascular shadow, and 28 lesions had drainage vein shadow adjacent to the lesions, 24 of which were located at the proximal end of the lesions. Fascial tail sign was found at the periphery of the lesions in 26 cases, of which 23 cases were located at the upper and lower ends of the lesions.Conclusion:FAVA is a complex vascular malformation with unclear pathological classification. MRI usually shows a solid mass in the superficial muscle layer of the lower limbs parallel to the long axis of the muscle, with cloud-like, band-like, and patchy hyperintensity on the background of high signal on T1WI, and dendritic, band-like, and cloud-like hypointensity on the background of high signal on T2WI-FS, which is helpful for the diagnosis of FAVA. Combined with the data of fascial tail sign, draining vein and clinical manifestations, the diagnosis of FAVA can be confirmed to a certain extent, which can provide reference and basis for clinical decision making.

ObjectiveTo investigate the value of ultrasonography as a diagnostic aid in differentiating intramuscular capillary-type hemangioma (ICTH) from fibro-adipose vascular anomaly (FAVA).MethodsA retrospective analysis was conducted of the clinical and ultrasound imaging data of 20 patients with ICTH and 45 patients with FAVA who were admitted to and pathologically confirmed in hospital between January 2013 and April 2023. The clinical and ultrasonographic appearances of the lesions in the two groups were compared and analyzed. A stepwise regression analysis was performed, and a joint diagnostic equation was constructed using the final variables selected. The receiver operating characteristic (ROC) curve and indicators, including sensitivity and specificity, were used to evaluate the efficacy of the joint diagnostic model.ResultsThe two groups of patients suffering from ICTH and FAVA presented a statistically significant difference (P< 0.05) in terms of ‘age’, ‘lesion size’, ‘fascial tail sign’, ‘presence of a fatty-tissue-like hyperecho around the lesion’, ‘blood flow’ and ‘presence of straight blood capillaries within the lesion’. Finally, the variables ‘fascial tail sign’ and ‘presence of straight blood capillaries within the lesion’ were selected to construct the model. The constructed joint diagnostic model had a sensitivity value of 70.0% (95% CI: 59.00–81.00), a specificity value of 98.0% (95% CI: 94.70–100.00) and a ROC curve value of 0.908, indicating the high efficacy of the combined diagnosis method.ConclusionsUltrasonography can be utilized to differentiate ICTH from FAVA, and the combined diagnosis method can further improve the technique’s diagnostic efficacy.

目的 观察儿童疣状血管瘤(VH)特征性超声、MRI及临床表现.方法 回顾性分析25例经术后病理证实的VH患儿,观察VH影像学及临床特征性表现.结果 25例VH患儿皮肤见不规则红色、青紫色斑块,斑块边缘皮肤呈暗红色,范围较广时呈"铠甲样"外观,周边见"卫星状"病灶.15例仅接受超声检查,体表疣状物多呈低回声、后方伴声影,深部皮下组织以高回声为主;其中10例病灶呈均匀高回声、4例高低混杂回声、1例均匀低回声;13例病灶边界不清,2例边界清晰;12例病灶内未见明显血流信号,3例见少量血流信号.10例仅接受MR检查;其中9例病灶T1WI呈低信号、脂肪抑制(FS)-T2WI呈不均匀稍高信号,1例病灶呈等TIWI信号、FS-T2WI高信号.结论 儿童VH皮肤病变多呈不规则红色、青紫色斑块,斑块边缘皮肤呈暗红色,可形成"铠甲样"外观,伴周边"卫星状"病灶;超声多表现为均匀高回声且边界不清;MRI多呈T1WI低信号、FS-T2WI不均匀稍高信号.

Objective:To investigate the differential performance of ultrasound between fibro-adipose vascular anomaly (FAVA) and venous malformations(VMs).Methods:From January 2015 to December 2020, the patients diagnosed with lower extremity FAVA by pathology in Henan Provincial People’s Hospital were enrolled as FAVA group. The patients diagnosed with lower extremity VMs by pathology were enrolled as the control group. The clinical and ultrasound imaging data were retrospectively analyzed. Through the single factor analysis of the two groups’data, the ultrasonic imaging indicators which may be valuable for distinguishing FAVA from VMs were screened. Further, the indicators valuable for differential diagnosis were determined by multi-factor Logistic regression analysis, and a multi-factor joint diagnosis model was constructed. The diagnostic efficiency of the joint diagnosis model was evaluated by the receiver operator characteristic curve (ROC curve), sensitivity, and specificity of the subjects.Results:A total of 20 patients with FAVA were involved, including 11 males and 9 females. The mean age was (18.1±12.2) years. Forty-six patients with VMs were involved, including 20 males and 26 females. The mean age was (19.9±13.6) years. Results of the single-factor analysis were differences in the lesion echo, fascial tail, blood flow, extrusion test, and posterior echo enhance characteristics between groups ( P<0.05). Multivariate analysis showed significant differences between groups in three aspects: fascial tail, extrusion test, and posterior echo enhancement ( P=0.001, 0.008, 0.007). The sensitivity and specificity of the multi-factor combined diagnosis model were 90.0% (95% CI: 68.3%-98.8%) and 93.5%(95% CI: 82.1%-98.6%), indicating high diagnostic efficiency. The ROC(AUC) area was 0.964(95% CI: 0.886-0.994), indicating high diagnostic efficiency. Conclusions:The ultrasonic imaging features of FAVA and VMs were different. The combined diagnosis of the fascial tail, compression test, and posterior echo enhancement has a higher auxiliary diagnostic value.

目的:探讨microRNA-192(miR-192)对于高侵袭性结肠癌RKO细胞的增殖、迁移和侵袭能力的影响.方法:将miR-192 mimics(miR-192)通过脂质体转染至RKO细胞,应用qRT-PCR检测细胞中miR-192表达情况,采用CCK-8实验和集落形成实验研究miR-192对结肠癌RKO细胞增殖的影响,采用细胞划痕实验和Transwell小室模型检测miR-192对结肠癌细胞迁移和侵袭能力的影响,应用Western blot检测miR-192对结肠癌RKO细胞内上皮性钙黏附蛋白(E-cadherin,E-cad)和血管内皮生长因子(vascular en-dothelial growth factor,VEGF)蛋白表达的影响.结果:CCK-8实验和集落形成实验结果显示:转染了miR-192的结肠癌RKO细胞,癌细胞增殖显著受到抑制.细胞划痕实验、迁移实验和细胞侵袭实验结果显示:RKO细胞转染miR-192后,迁移和侵袭的细胞数量显著减少,差异有统计学意义(P<0.05).Western blot结果显示:结肠癌细胞转染miR-192后可升高E-cad蛋白的表达,并且降低VEGF蛋白的表达.结论:转染miR-192可抑制结肠癌RKO细胞增殖,降低其迁移和侵袭能力,其可能的机制是升高结肠癌RKO细胞E-cad的表达,并且降低VEGF的表达.miR-192可能作为肿瘤诊断及治疗的新靶点.

血管肌纤维母细胞瘤(angiomyofibroblastoma,AMF)是一种极少见的特殊类型的良性间叶性肿瘤,我们回顾性分析10例AMF,并结合相关文献,探讨该类肿瘤的临床病理特点,旨在加深临床和病理医师对该病的认识.

目的 探讨金雀异黄素(genistein,GEN)诱导乳腺癌MDA-MB-231细胞凋亡的分子机制.方法 用0、5、10、20 μmol/L GEN处理MDA-MB-231 细胞24 h.采用CCK-8、Hoechst 33342染色和流式细胞仪测定不同浓度GEN对 MDA-MB-231细胞增殖和凋亡的影响.采用Western blotting检测不同浓度GEN处理前后MDA-MB-231细胞中Fas相关死亡域蛋白(FADD)、活性半胱天冬酶8(cleaved caspase-8)、Fas、FasL蛋白表达水平.采用实时RT-PCR分析不同浓度GEN处理前后MDA-MB-231细胞中Fas、FasL基因表达水平.多组均数比较采用方差齐性检验后进行单因素方差分析.结果 在GEN作用24 h后,0、5、10和20 μmol/L组对MDA-MB-231细胞增殖的抑制率分别为(3.00±1.41)%、(14.02±1.57)%、(27.5±1.52)%、(48.90±1.44)%.与0 μmol/L组相比,5、10和20 μmol/L组呈浓度依赖性增加(F=528.119,P=0.000).两两比较显示:各浓度组之间差异均有统计学意义(P<0.05).0、5、10和20 μmol/L组诱导MDA-MB-231细胞的早期凋亡率分别为(3.40±0.40)%、(9.34±1.34)%、(19.26±0.93)%、(27.41±1.12)%.与0 μmol/L组相比,5、10和20 μmol/L组呈浓度依赖性增加(F=379.573,P=0.000).两两比较显示:各浓度组之间差异均有统计学意义(P<0.05).Western blotting结果显示,与0 μmol/L组相比,其他浓度组经GEN处理的MDA-MB-231细胞FADD、cleaved caspase-8、FasL蛋白表达升高(F=368.621、456.744、419.129,P均=0.000),Fas蛋白表达差异无统计学意义(F=0.800,P=0.528);与10(μmol/L组相比,20 μmol/L组FasL蛋白表达降低有统计学意义(F=92.235,P=0.001).实时RT-PCR结果显示,与0 μmol/L组相比,其他浓度组经GEN处理的MDA-MB-231细胞FasL mRNA表达升高(F=646.983,P=0.000),Fas mRNA表达差异无统计学意义(F=1.556,P=0.274);与10 μmol/L组相比,20 μmol/L组FasL mRNA表达降低有统计学意义(F=52.562,P=0.020).结论 GEN通过上调Fas/FasL途径中FasL基因表达诱导乳腺癌MDA-MB-231细胞凋亡.

Purpose To investigate the ALDH1,HIF-1α and VHL expression and their impact on the prognosis of patients with breast cancer.Methods Immunohistochemistry(SP method) was used to detect the stem cell marker ALDH1,VHL and HIF-1α expression in 15 cases of normal breast tissue,30 cases of breast hyperplasia and 40 cases of breast invasive ductal carcinoma.Results Positive expression rate of ALDH1 HIF-1α and VHL among normal tissue,breast hyperplasia,and breast invasive ductal carcinoma was statistically significant(P 0.05).Spearman rank correlation analysis showed that the VHL and HIF-1α,tumor grade,clinical stage,tumor size,lymph node metastasis were negatively correlated,in which HIF-1α and tumor grade,clinical stage,tumor size,lymph node metastasis were positively correlated(P 0.05).In addition,the expression of HIF-1α and VHL was associated with ALDH1 in breast cancer(r = 0.976,P = 0.001;r = 0.971,P = 0.001;respectively).Conclusion Abnormal expression of ALDH1,HIF-1α and VHL exists in breast cancer,which suggests that they may play an important role in the progression,invasion and metastasis.VHL / HIF-1α signaling pathway may be related to cancer stem cell growth and differentiation.

OBJECTIVE:To investigate the influence of down-regulating Smoothened (SMO) gene expression through short hairpin RNA (shRNA) on the proliferation of breast cancer stem cells.METHODS:Human SMO shRNA was designed, synthesized chemically, and transfected into MCF-7 cells to down-regulate SMO gene. By using G418, stable cells with down-regulated SMO were selected. In vitro proliferation of these cells was measured by CCK8 assay. The proportion of CD44(+)/CD24(-) cells was detected by flow cytometry and the mammospheres formation was determined by suspension sphere culture. The expression of SMO, GLI1 and Oct4 was detected by Western blot. In vivo, the volume of tumor was measured every 3 days and the expression of SMO, GLI1 and Oct4 detected by Western blot.RESULTS:In vitro, the cells were transfected with SMO-shRNA and selected by G418 after 21 days. SMO-shRNA effectively down-regulated the expression of SMO gene and protein, and inhibited the proliferation of MCF-7 and markedly reduced the proportion of CD44(+)/CD24(-) cells and mammospheres. In vivo, SMO-shRNA treatment of MCF-7 significantly inhibited the volume of tumor. The positive rate of SMO in negative control and SMO-shRNA group was 5/5 and 2/5, respectively. The expression of SMO, GLI1 and Oct4 in different groups were 0.72 ± 0.17 and 0.21 ± 0.09, 1.21 ± 0.21 and 0.47 ± 0.12, 0.83 ± 0.13 and 0.25 ± 0.07. SMO, GLI1 and Oct4 down-regulation significantly suppressed at protein levels (P < 0.05).CONCLUSION:The shRNA by chemical synthesis can effectively down-regulate SMO gene expression and inhibit the proliferation of breast cancer stem cells.

INTRODUCTION:The existence of breast cancer stem-like cells (BCSCs) has profound implications for cancer prevention. Genistein, a predominant isoflavone found in soy products, has multiple robust anti-tumor effects in various cancers, especially in the breast and prostate cancer. In this study, we aimed to evaluate genistein inhibition of BCSCs and its potential mechanism by culturing MCF-7 breast cancer cells and implanting these cells into nude mice.METHODS:Cell counting, colony formation and cell apoptosis analysis were used to evaluate the effect of genistein on breast cancer cells’ growth, proliferation and apoptosis. We then used mammosphere formation assay and CD44CD24 staining to evaluate the effect of genistein on BCSCs in vitro. A nude mice xenograft model was employed to determine whether genistein could target BCSCs in vivo, as assessed by real-time polymerase chain reaction (PCR) and immunohistochemical staining. The potential mechanism was investigated utilizing real-time PCR, western blotting analysis and immunohistochemical staining.RESULTS:Genistein inhibited the MCF-7 breast cancer cells’ growth and proliferation and promoted apoptosis. Both in vitro and in vivo genistein decreased breast cancer stem cells, and inhibited breast cancer stem-like cells through down-regulation of the Hedgehog-Gli1 Signaling Pathway.CONCLUSIONS:We demonstrated for the first time that genistein inhibits BCSCs by down-regulating Hedgehog-Gli1 signaling pathway. These findings provide support and rationale for investigating the clinical application of genistein in treating breast cancer, and specifically by targeting breast cancer stem cells.

Objective To explore the molecular mechanism of genistein-induced apoptosis in breast cancer MDA-MB-231 cells. Methods MDA-MB-231 cells were treated with 0,5,10,20 μmol / L genistein for 24 h,respectively. CCK-8,Hoechst 33342 staining and flow cytometry were used to determine the effects of genistein on proliferation and apoptosis of MDA-MB-231 cells. The expressions of FADD,cleaved caspase-8, Fas,FasL in the protein level were measured by Western blotting. The expressions of Fas and FasL in the mRNA level were detected by real-time RT-PCR. The one-way ANOVA was conducted after homogeneity for variance was tested. Results The cell proliferation inhibition rate of MDA-MB-231 cells were( 3. 00 ± 1. 41) %,( 14. 02 ± 1. 57) %,( 27. 5 ± 1. 52) %,( 48. 90 ± 1. 44) % after the treatment of 0,5,10, 20 μmol / L genistein for 24 h respectively. Compared with 0 μmol / L group,the inhibition rate was increased in a concentration-dependent manner in the other 3 groups( F = 528. 119,P = 0. 000). The pairwise comparison showed the difference was statistically significant( all P < 0. 05). The early apoptotic rate of MDA-MB-231 cells were( 3. 40 ± 0. 40) %,( 9. 34 ± 1. 34) %,( 19. 26 ± 0. 93) %,( 27. 41 ± 1. 12) % in each concentration group, respectively,after genistein treatment for 24 h. Compared with 0 μmol / L group,the early apoptotic rate was increased in a concentration-dependent manner in the other 3 groups( F = 379. 573,P = 0. 000). The pairwise comparison showed the difference was statistically significant( all P < 0. 05). Western blotting showed that genistein increased the protein expression levels of FADD,cleaved caspase-8 and FasL( F = 368. 621,456. 744, 419. 129; all P = 0. 000),with no alteration in Fas protein level( F = 0. 800,P = 0. 528). Compared with 10 μmol / L group,FasL protein expression decreased in 20 μmol / L group( F = 92. 235,P = 0. 001). Real-time RT-PCR showed that genistein increased the mRNA expression of FasL( F = 646. 983,P = 0. 000),with alteration in Fas mRNA level( F = 1. 556,P = 0. 274). Compared with 10 μmol / L group,FasL mRNA expression also decreased in 20 μmol / L group( F = 52. 562,P = 0. 020). Conclusion Genistein induces apoptosis of breast cancer MDA-MB-231 cells by up-regulating FasL gene expression in Fas / FasL pathway.

Objective To investigate the effects of genistein on proliferation,invasion and metastasis of MDA-MB-231 breast cancer cells.Methods The proliferation of MDA-MB-231 breast cancer cells was measured by CCK 8 assay and flow cytometry assay. Transwell assay and Western blot assay were used to evaluate the migration and invasion ability of MDA-MB-231 breast cancer cells.Results 24h after genistein treatment,CCK-8 assay showed that the proliferation of breast cancer cells was inhibited with an IC50 value of 18.7 μmol/L Cell cycle analysis by flow cytometry as say confirmed that the proportion of cells in S phase decreased significantly after genistein treatment(24h). The number of migrating cells in tran swell assay was sharply reduced. The invasion of MDA-MB-231 cells was inhibited compared with the blank control group(P0.05). Western blot assay revealed that MMP-2 and VEGF decreased significantly by 10 and 20 μmol/L genistein.Conclusion Genistein could effectively inhibit the growth and invasionability of MDA-MB-231 breast cancer cells in a dose dependent pattern in vitro,and the reduction of MMP-2 and VEGFex pression may play animportant role in this process.

饮食中的genistein在降低亚洲人乳腺癌的发病率上有很重要的作用.最新研究表明genistein抑制乳腺癌细胞生长是通过调节细胞的增殖、分化和凋亡等相关信号转导通路来完成的.乳腺癌干细胞可能是乳腺癌发生耐药、复发及转移的根源.近年来发现乳腺癌干细胞相关信号转导通路复杂且涉及面广泛,其中蛋白酪氨酸激酶(protein tyrosine kinases,PTKs)信号转导通路在癌干细胞中的异常过表达,是引起癌基因转化、肿瘤生长的重要因素之一.因此,Genistein和乳腺癌干细胞相关的PTKs信号转导通路之间的关系也越来越受到关注,对寻找更为有效的乳腺癌靶向治疗具有重要意义.