Genomic characterization reveals distinct mutational landscapes and therapeutic implications between different molecular subtypes of triple-negative breast cancer.

Triple-negative breast cancer (TNBC) has high heterogeneity, poor prognosis, and limited treatment success. Recently, an immunohistochemistry-based surrogate classification for the "Fudan University Shanghai Cancer Center (FUSCC) subtyping" has been developed and is considered more suitable for clinical application. Seventy-one paraffin-embedded sections of surgically resected TNBC were classified into four molecular subtypes using the IHC-based surrogate classification. Genomic analysis was performed by targeted next-generation sequencing and the specificity of the subtypes was explored by bioinformatics, including survival analysis, multivariate Cox regression, pathway enrichment, Pyclone analysis, mutational signature analysis and PHIAL analysis. AKT1 and BRCA1 mutations were identified as independent prognostic factors in TNBC. TNBC molecular subtypes encompass distinct genomic landscapes that show specific heterogeneities. The luminal androgen receptor (LAR) subtype was associated with mutations in PIK3CA and PI3K pathways, which are potentially sensitive to PI3K pathway inhibitors. The basal-like immune-suppressed (BLIS) subtype was characterized by high genomic instability and the specific possession of signature 19 while patients in the immunomodulatory (IM) subtype belonged to the PD-L1 ≥ 1% subgroup with enrichment in Notch signaling, suggesting a possible benefit of immune checkpoint inhibitors and Notch inhibitors. Moreover, mesenchymal-like (MES) tumors displayed enrichment in the receptor tyrosine kinase (RTK)-RAS pathway and potential sensitivity to RTK pathway inhibitors. The findings suggest potential treatment targets and prognostic factors, indicating the possibility of TNBC stratified therapy in the future.

CDKN2A promoter methylation enhances self-renewal of glioblastoma stem cells and confers resistance to carmustine.

BACKGROUND: Glioblastoma, a highly aggressive form of brain cancer, poses significant challenges due to its resistance to therapy and high recurrence rates. This study aimed to investigate the expression and functional implications of CDKN2A, a key tumor suppressor gene, in glioblastoma cells, building upon the existing background of knowledge in this field. METHOD: Quantitative reverse transcription PCR (qRT-PCR) analysis was performed to evaluate CDKN2A expression in U87 glioblastoma cells compared to normal human astrocytes (NHA). CDKN2A expression levels were manipulated using small interfering RNA (siRNA) and CDKN2A overexpression vector. Cell viability assays and carmustine sensitivity tests were conducted to assess the impact of CDKN2A modulation on glioblastoma cell viability and drug response. Sphere formation assays and western blot analysis were performed to investigate the role of CDKN2A in glioblastoma stem cell (GSC) self-renewal and pluripotency marker expression. Additionally, methylation-specific PCR (MSP) assays and demethylation treatment were employed to elucidate the mechanism of CDKN2A downregulation in U87 cells. RESULT: CDKN2A expression was significantly reduced in glioblastoma cells compared to NHA. CDKN2A overexpression resulted in decreased cell viability and enhanced sensitivity to carmustine treatment. CDKN2A inhibition promoted self-renewal capacity and increased pluripotency marker expression in U87 cells. CDKN2A upregulation led to elevated protein levels of p16INK4a, p14ARF, P53, and P21, which are involved in cell cycle regulation. CDKN2A downregulation in U87 cells was associated with high promoter methylation, which was reversed by treatment with a demethylating agent. CONCLUSION: Our findings demonstrate that CDKN2A downregulation in glioblastoma cells is associated with decreased cell viability, enhanced drug resistance, increased self-renewal capacity, and altered expression of pluripotency markers. The observed CDKN2A expression changes are mediated by promoter methylation. These results highlight the potential role of CDKN2A as a therapeutic target and prognostic marker in glioblastoma.

JAK/STAT3 Signaling Activation Related to Distinct Clinicopathologic Features in Systemic ALK - Anaplastic Large Cell Lymphomas : New Insights into Their Heterogeneity.

Systemic anaplastic lymphoma kinase (ALK)-negative anaplastic large cell lymphoma (ALCL) is a group of heterogenous CD30 + T-cell non-Hodgkin lymphomas. Previous studies have highlighted the importance of JAK/STAT3 signaling activation in the molecular pathogenesis of ALK - ALCLs. In the present study, we aimed to establish a potential relationship between JAK/STAT3 signaling activation and clinicopathologic features in ALK - ALCLs, and further recognize the heterogenous nature of these neoplasms. Immunohistochemistry staining of the phosphorylated-STAT3 (p-STAT3) and dual-specificity protein phosphatase 22 ( DUSP22 ) gene rearrangement analysis were performed. Forty-five cases of ALK - ALCL were divided into 3 groups, including 9 DUSP22 -rearranged ALCLs, 21 p-STAT3 + double-negative (DN) ALCLs (both ALK and DUSP22 rearrangement negative), and 15 p-STAT3 - DN-ALCLs. Morphologically, p-STAT3 + DN-ALCLs exhibited sheet-like neoplastic cells and sometimes showed large pleomorphic cells scattered in a lymphocyte-rich background more frequently than those in other ALK - ALCLs subtypes. Phenotypically, the p-STAT3 + DN-ALCLs frequently expressed cytotoxic molecules, epithelial membrane antigen, and programmed death-ligand 1, whereas CD3 and CD5 expression was not observed. Clinically, patients with p-STAT3 + DN-ALCLs had a better prognosis than those with p-STAT3 - DN-ALCLs. These observations suggest that p-STAT3 + DN-ALCLs represent a distinct subtype of ALK - ALCLs. Identifying ALK - ALCL subtypes by using p-STAT3 staining and DUSP22 rearrangement is a promising approach that may contribute to risk stratification and better treatment decisions in the future clinical practice.

Ovarian mucinous tumor with mural nodules of anaplastic carcinoma: Three case reports.

BACKGROUND: Anaplastic carcinoma mural nodules in ovarian mucinous tumors are very rare. This study aimed to report the morphological characteristics, molecular detection results, clinical treatment and prognosis of three ovarian mucinous tumors with mural nodules of anaplastic carcinoma. CASE SUMMARY: The pathomorphological features, molecular detection results, clinical treatment and prognosis of anaplastic carcinoma mural nodules were described in three cases. In case 1, sarcoma-like mural nodules (SLMNs) coexisted with anaplastic carcinoma mural nodules. No mutation was found in mucinous tumors. KRAS mutation was found in anaplastic carcinoma nodules and heterotypic cells were found in SLMNs. In case 2, KRAS mutation occurred in the mucinous epithelium and BRAF mutation occurred in mural nodules. In case 3, both mural nodules and mucinous tumors had the same KRAS mutation and a morphological transition between them was observed. All three patients died within 2 years, whether receiving chemotherapy or not. CONCLUSION: Anaplastic carcinoma mural nodules may develop from dedifferentiation of mucinous tumors or are unrelated to mucinous tumors.

Subclassifying triple-negative breast cancers and its potential clinical utility.

The molecular subtyping of triple-negative breast cancer (TNBC) is critical to guiding individualized patient treatment. In this study, we sought to characterize the clinicopathologic features of TNBC subtypes and to identify correlates of patient survival in an effort to provide a robust foundation for treatment planning. We additionally assessed PD-L1 expression in Chinese TNBC patients and evaluated the relationship between such expression and immunotherapeutic treatment outcomes. Based on analyses of histologic characteristics including apocrine differentiation, tumor-infiltrating lymphocytes, and metaplastic features, we selected immunohistochemical (IHC) markers including CD8, FOXC1, and AR for use in classifying TNBC cases. Associations between these subtypes and a range of clinicopathologic characteristics were evaluated. We classified a cohort of 93 TNBC patients into individuals with luminal androgen receptor (LAR), immunomodulatory (IM), basal-like immune-suppressed (BLIS), and mesenchymal (MES) tumor subtypes (23, 24, 39, and 7 cases, respectively). PD-L1 positivity was observed in 49.6% of cases and was more common in individuals with IM subtype disease. Mismatch repair deficiency (dMMR) was observed in just one patient. Significant differences in histologic grade, pT stage, lymphocyte distribution patterns, large scarring areas without cells in tumor of central (central scar), and PD-L1, P53, and Rb status were observed among these TNBC subtypes, whereas no such differences were observed with respect to age, invasion pattern, or pN stage. Rates of disease progression were higher at the 40-50 month follow-up time point, but there were no significant differences in recurrence-free survival or breast cancer-specific survival among these subtypes. IHC markers associated with clinicopathologic characteristics represent a powerful approach to TNBC molecular typing, providing a foundation for precision patient treatment. PD-L1 expression may represent a relevant factor in TNBC patient immunotherapeutic treatment planning, whereas dMMR is not likely to be of substantial value when evaluating immunotherapeutic efficacy in these patients.

Secretory breast carcinoma in a female adult with liver metastsis: a case report and literature review.

BACKGROUND: Secretory breast carcinoma is an uncommon subset of breast cancer that usually has a favorable outcome. Although initially described in children, it also occurs in adults where it may metastasize, possibly resulting in death. To date, only 20 cases of secretory breast carcinoma with distant metastases have been described. CASE PRESENTATION: A 42-year-old female presented with liver metastasis after modified radical mastectomy of the left breast in 2008 at 34 years of age. The liver metastasis was morphologically similar to the primary tumor. Pan-TRK and Fluorescence in situ hybridization showed a rearrangement in the ETV6 gene. She subsequently underwent adjuvant chemotherapy with a fatal outcome. CONCLUSIONS: Although secretory breast carcinoma is usually associated with favorable outcomes, our study and reviews provide a novel insight into the genetic spectrum and treatment of secretory breast carcinoma showing reduced expression of hormone receptors, abnormal genomic profiles, and possible poor prognosis. Targeted therapy may curb clinically aggressive cases. Additional molecular investigations are needed to determine the links between specific mutations and poor prognosis.

Clinical-Pathologic Analysis of Breast Cancer With PIK3CA Mutations in Chinese Women.

PURPOSE: Mutations of PIK3CA have recently been shown to play an important role in the pathogenesis and progression of breast neoplasms. The prevalence of PIK3CA in Chinese breast cancer patients may be underestimated. Therefore, we investigated the distribution of somatic PIK3CA mutation in Chinese breast cancer patients and explored their role in tumor phenotypes. METHODS: Mutational analysis of PIK3CA was done in 113 primary breast cancers of Chinese women used Amplification refractory mutation system (ARMS). The relationship of PIK3CA mutations with several clinicopathologic characteristics was analyzed. RESULTS: PIK3CA gene mutation was identified in 43(38.05%) cases and has a more significant difference between exon 9 and 20. HER2 gene amplification was 32.6% in 43 cases of PIK3CA mutation, but 37.1% in 70 cases of non-mutation (χ2 = 0.245, P > 0.05). There was no significant correlation of the age distribution, lymph node status, histological tumor grading, ER and/or PR and P53 between 2 groups (P > 0.05). CONCLUSION: A high frequency of somatic PIK3CA mutation was detected in Chinese breast cancer patients, especially in exon 20. The relationship between PIK3CA gene mutation and clinical pathological features of breast cancer needs to be further studied in a large series of patients. PIK3CA mutations seem to have the potential to be used in target treatment and as an indicator of prognosis.

[Significance of microRNA-16 and bcl-2 expression in T lymphoblastic lymphoma/leukemia and its relation with prognosis].

OBJECTIVE: To study the expression of miR-16 and bcl-2 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) and its relationship to prognosis. METHODS: 70 cases of T-LBL/ALL with follow-up data were studied by using immunohistochemical EnVision method for CD1a, CD3, cCD3, CD7, CD10, CD20, CD23, CD34, CD43, CD45RO, CD99, TDT, MPO, bcl-2 and Ki-67. The expression levels of miR-16 were examined by TaqMan real-time polymerase chain reaction (RT-PCR). Thirty cases of reactive lymph node were selected as control. RESULTS: Among the 70 cases of T-LBL/ALL, the percentages of tumor cells expression of TDT, CD99, CD3, CD7, CD10, CD34, CD1a, cCD3, bcl-2, CD45RO and CD43 were 94.3% (66/70), 94.3% (66/70), 68.6% (48/70), 92.9% (65/70), 32.9% (23/70), 24.3% (17/70), 40.0% (28/70), 51.4% (36/70), 34.3% (24/70), 37.1% (26/70), and 48.6% (34/70). Separately, while tumor cells expression of MPO, CD20 and CD23 was all negative. A figure of Ki-67 expression > 80% was found in 24 cases and ≤ 80% in 46 cases. The expression of miR-16 was up-regulated in T-LBL/ALL, and it was 5.07 times of the reactive lymph node(P = 0.001). The high expression group of miR-16 was significantly correlated with longer over survival (P = 0.041). The prognosis of negative bcl-2 group was better than bcl-2 positive one(P = 0.904). The relationship of miR-16 and bcl-2 was significant(P = 0.042,χ(2) = 4.147). Survival multivariate COX proportional hazard regression analysis revealed that the low expression of miR-16 might be a independent poor prognosis factor (P = 0.049). CONCLUSIONS: While the high expression group of miR-16 has longer OS than that in low expression group. The prognosis of bcl-2 negative was better than bcl-2 positive. miR-16 may be a independent prognosis factor. The relationship of miR-16 and bcl-2 might suggested that gene regulation may be influenced by them.

[Correlation of Bcl-2 with immunological subtype and prognosis in diffuse large B-cell lymphoma.].

OBJECTIVE: To investigate the Bcl-2 protein and gene expression in diffuse large B-cell lymphoma (DLBCL), and analyze its correlation with immunosubtype and prognosis. METHODS: Seventy-three cases of DLBCL were performed immunohistochemistry analysis with a panel of antibodies CD3, CD10, CD20, Bcl-6, Bcl-2 and MUM-1, and classified into germinal center B-cell (GCB) type and non-GCB type. Fluorescence in situ hybridization (FISH) was employed to detect bcl-2 gene expression in 57 cases with chromosome translocation t (14;18). RESULTS: The percentages of tumor cells expressed CD10, Bcl-6, MUM-1 and Bcl-2 were 15.1%, 38.4%, 71.2% and 79.2%, respectively. 16 cases (21.9%) were GCB type and the rest (78.1%) were non-GCB type. 16 of 57 cases (28.1%) were t (14; 18), including 5 of GCB type (31.2%) and 11 of non-GCB type (68.2%). The expression of Bcl-2 protein was correlated with immunological subtype (P = 0.035), but not with survival time (P = 0.253). Between the t(14;18) positive and negtive groupes, there was significant difference for survival time (P = 0.022), but no difference for immunological subtype (P = 0.340). There was no correlation between Bcl-2 protein and t(14;18). CONCLUSIONS: GCB type DBLBCL with expression of Bcl-2 protein had a poor prognosis. t(14; 18) positive BLBCL had poor prognosis. The expression of Bcl-2 protein and t(14; 18) are usually discordant.

[State of chromosome 3q27 and different subtypes of diffuse large B-cell lymphoma and their prognostic correlation].

OBJECTIVE: To study the immunophenotypic and genetic features of diffuse large B-cell lymphomas (DLBCLs), and their relationship to prognosis. METHODS: Seventy-three cases of DLBCLs with follow-up data were studied by using immunohistochemistry for CD3, CD10, CD20, bcl-6 and MUM-1. The DLBCLs were classified into germinal center B cell-like (GCB) and non-germinal center B cell-like (non-GCB) subtypes according to Hans' algorithm. Fluorescence in-situ hybridization (FISH) for bcl-6 gene expression (located on chromosome 3q27) was performed on paraffin-embedded tissues of 54 cases. RESULTS: In the 73 cases studied, 16 cases (21.9%) belonged to GCB subtype and 57 cases (78.1%) belonged to non-GCB subtype. Breakage of 3q27 was detected in 11 of the 54 cases (20.4%) and proliferation was detected in 14 cases (25.9%). The five-year overall survival rate of GCB subtype was significantly higher than that of non-GCB subtype (78% versus 40%, P = 0.011). The bcl-6-positive cases had a better clinical outcome than that of the bcl-6-negative cases (P = 0.041). Breakage of 3q27 predicted a worse overall survival. CONCLUSIONS: The current study shows that the prognosis of GCB subtype of DLBCLs is better than that of non-GCB subtype. The expression of bcl-6 protein predicts a better clinical outcome, while the breakage of 3q27 predicts a worse overall survival.

[Study on the origin of H/RS cell and their biological behavior in Hodgkin lymphoma by using multiple mark techniques].

OBJECTIVE: To investigate the apoptosis-related genes and protein expression patterns in relation to classical Hodgkin lymphomas (CHL) and the origin of H/RS cell. METHODS: Sixty-two cases of CHL were retrieved from Shanxi Tumor Hospital files. An ABC method was used to detect the expression of bcl-2, CD3, CD20, CD30, CD15 and CD10, a double immunohistochemical method to study the H/RS cells P53 expression, a double immunohistochemical ABC-DNA end labeling technique to detect the apoptosis, a double immunohistochemical ABC- in situ hybridization technique to detect the expression of kappa mRNA and lambda mRNA, and a multiple mark techniques to detect the distribution of background non-neoplastic T and B cells. RESULT: Of 62 CHL, 14 (22.58%) were p53 positive and 35 (56.45%) bcl-2 positive. Apoptosis was found in the background non-neoplastic cells in all of the cases, but in H/RS cells in only 10 of 62 cases. There was a significant reverse correlation between bcl-2 expression and apoptosis in H/RS cells (P = 0.02). CD30 positive H/RS cells were observed in all cases, whereas CD15 positive in only 41 cases, and CD20 positive in 8 cases. None was positive for CD3, MPO, bcl-6, CD10, kappa RNA and lambda RNA in H/RS cells. The H/RS cells were surrounded by non-neoplastic T cells looked like a rosette and the outer periphery was B cells. CONCLUSIONS: The H/RS cell of classical Hodgkin lymphoma has a great variety of B lineage markers. The characteristic distributions of T, B and H/RS cells may serve as a reference for the diagnosis. Multiple marker technique is able to highlight the critical cells, and facilitate the study of H/RS cells. Abnormal expression of P53 may not play a major role in CHL. Over expression of bcl-2 may be linked to blockage of apoptosis in CHL.