Rare germline mutation and MSH2-&MSH6 + expression in a double primary carcinoma of colorectal carcinoma and endometrial carcinoma: a case report.

BACKGROUND: Multiple primary malignancies are rare in cancer patients, and risk factors may include genetics, viral infection, smoking, radiation, and other environmental factors. Lynch syndrome (LS) is the most prevalent form of hereditary predisposition to double primary colorectal and endometrial cancer in females. LS, also known as hereditary nonpolyposis colorectal cancer (HNPCC), is a common autosomal dominant condition. Pathogenic germline variants in the DNA mismatch repair (MMR) genes, namely MLH1, MSH2, MSH6, and PMS2, and less frequently, deletions in the 3' end of EPCAM cause LS. It manifested itself as loss of MMR nuclear tumor staining (MMR protein deficient, dMMR). CASE PRESENTATION: This case study describes a double primary carcinoma in a 49-year-old female. In June 2022, the patient was diagnosed with highly to moderately differentiated endometrioid adenocarcinoma. The patient's mother died of esophageal cancer at age 50, and the father died of undefined reasons at age 70. Immunohistochemical stainings found ER (++), PR (++), P53 (+), MSH2 (-), MSH6 (+), MLH1 (+), and PMS2 (+). MMR gene sequencing was performed on endometrial tumor and peripheral blood samples from this patient. The patient carried two pathogenic somatic mutations in the endometrial tumor, MSH6 c.3261dupC (p.Phe1088LeufsTer5) and MSH2 c.445_448dup (p.Val150fs), in addition to a rare germline mutation MSH6 c.133G > C (p.Gly45Arg). Two years ago, the patient was diagnosed with moderately differentiated adenocarcinoma in the left-half colon. Immunohistochemical stainings found MSH2(-), MSH6(+), MLH1(+), and PMS2(+) (data not shown). CONCLUSIONS: In the case of a patient with double primary EC and CRC, a careful evaluation of the IHC and the genetic data was presented. The patient carried rare compound heterozygous variants, a germline missense mutation, and a somatic frameshift mutation of MSH6, combined with a novel somatic null variant of MSH2. Our study broadened the variant spectrum of double primary cancer and provided insight into the molecular basis for abnormal MSH2 protein loss and double primary carcinoma.

Iterative multiple instance learning for weakly annotated whole slide image classification.

Objective.Whole slide images (WSIs) play a crucial role in histopathological analysis. The extremely high resolution of WSIs makes it laborious to obtain fine-grade annotations. Hence, classifying WSIs with only slide-level labels is often cast as a multiple instance learning (MIL) problem where a WSI is regarded as a bag and tiled into patches that are regarded as instances. The purpose of this study is to develop a novel MIL method for classifying WSIs with only slide-level labels in histopathology analysis.Approach.We propose a novel iterative MIL (IMIL) method for WSI classification where instance representations and bag representations are learned collaboratively. In particular, IMIL iteratively finetune the feature extractor with selected instances and corresponding pseudo labels generated by attention-based MIL pooling. Additionally, three procedures for robust training of IMIL are adopted: (1) the feature extractor is initialized by utilizing self-supervised learning methods on all instances, (2) samples for finetuning the feature extractor are selected according to the attention scores, and (3) a confidence-aware loss is applied for finetuning the feature extractor.Main results.Our proposed IMIL-SimCLR archives the optimal classification performance on Camelyon16 and KingMed-Lung. Compared with the baseline method CLAM, IMIL-SimCLR significantly outperforms it by 3.71% higher average area under curve (AUC) on Camelyon16 and 4.25% higher average AUC on KingMed-Lung. Additionally, our proposed IMIL-ImageNet achieve the optimal classification performance on TCGA-Lung with the average AUC of 96.55% and the accuracy of 96.76%, which significantly outperforms the baseline method CLAM by 1.65% higher average AUC and 2.09% higher average accuracy respectively.Significance.Experimental results on a public lymph node metastasis dataset, a public lung cancer diagnosis dataset and an in-house lung cancer diagnosis datasets show the effectiveness of our proposed IMIL method across different WSI classification tasks compared with other state-of-the-art MIL methods.

The prognostic value of Tiam1 correlates with its roles in epithelial-mesenchymal transition progression and angiogenesis in lung adenocarcinoma.

BACKGROUND: Tiam1 has been identified as an oncogene and acts as an activator of GTPase Rac. Tiam1 was reported to be a promoter of cancer progression in various cancer types, while in lung adenocarcinoma, its mechanism of action is poorly understood. MATERIALS AND METHODS: Immunohistochemistry staining and Western blot assay were used to determine Tiam1 expression in lung adenocarcinoma tissues, and its association with prognosis was determined by statistical analysis. We depleted Tiam1 in both A549 and H1975 cancer cell lines. Carboxyfluorescein diacetate succinimidyl ester staining and colony formation assays were used to evaluate its impact on cell proliferation ability after depletion. Transwell migration assay and wound healing assays were performed to determine its impact on migration ability of both cell lines. Western blot assay and immunofluorescence staining were used to analyze the association between Tiam1 and epithelial-mesenchymal transition (EMT) progression. Tube formation assay and vasculogenic mimicry assay were used to show the impact of Tiam1 depletion on cancer angiogenesis. RESULTS: In this study, we demonstrated that Tiam1 overexpression in lung adenocarcinoma was significantly associated with advanced tumor grade and poor prognosis. In vitro assays indicated that Tiam1 depletion significantly inhibited cell proliferation, colony formation, and migration capacities in A549 and H1975 cells. Further investigations revealed that Tiam1 plays an important role in EMT program enhancement, angiogenesis, and accelerated tumor progression. Notably, Tiam1 depletion in cancer cells strongly inhibited human umbilical vein endothelial cell angiogenesis and vasculogenic mimicry capacities of both cancer cell lines. CONCLUSION: Tiam1 overexpression is associated with lung adenocarcinoma progression and may indicate poor prognosis. Tiam1 accelerated tumor progression due to EMT and angiogenesis enhancement. Our data may provide a novel therapeutic target for lung adenocarcinoma.

Elevated expression of Tiam1 is associated with poor prognosis and promotes tumor progression in pancreatic cancer.

OBJECTIVE: T-cell lymphoma invasion and metastasis inducing factor 1 (Tiam1) is known to be involved in tumor progression. However, its molecular roles and mechanism in pancreatic ductal adenocarcinoma (PDAC) remain unclear. The purpose of this study is to determine Tiam1 expression levels and investigate its underlying molecular mechanism in PDAC. MATERIALS AND METHODS: Tiam1 protein expression levels in PDAC tissues were examined using immunohistochemistry. Tiam1 expression was confirmed in pancreatic cancer (PC) cells by Western blot and immunofluorescence staining. Tiam1-silenced PC cells were created using short interfering RNA. Subsequently, colony formation, scratch, and migration and invasion assays were carried out to explore the molecular mechanisms of Tiam1 in PC cells. RESULTS: The results indicated that Tiam1 expression was significantly higher in PDAC tissues than in paired non-tumor tissues, and overexpression of Tiam1 was significantly correlated with histological grade (P=0.040) and lymph node metastasis (P=0.031) in PDAC. The PDAC patients with high Tiam1 expression had significantly lower 5-year overall survival than patients with low Tiam1 expression. More importantly, univariate and multivariate analysis suggested that Tiam1 expression, along with lymph node metastasis, is a significant independent prognostic factor for patients with PDAC. Furthermore, we also demonstrated that the downregulation of Tiam1 was associated with decreased cell proliferation and reduced migratory and invasive capability. CONCLUSION: High expression of Tiam1 plays a significant role in the progression of PDAC and may be a potential biomarker of poor prognosis as well as a therapeutic target.

Upregulation of Tiam1 contributes to cervical cancer disease progression and indicates poor survival outcome.

T lymphoma invasion and metastasis 1 (Tiam1), a guanine nucleotide exchange factor, is involved in the tumorigenesis of a number of malignancies. This study was aimed to explore the role of Tiam1 in cervical cancer progression, and evaluate the prognostic values. Tiam1 protein expression levels were detected by immunohistochemical (IHC) staining in 174 cervical cancer tissues, 92 of CINs (cervical intraepithelial neoplasia) and 32 of normal cervical epithelia tissues. Clinicopathological parameters and overall survival data were collected and compared between different Tiam1 statuses. The role of Tiam1 in cervical cancer proliferation, migration and angiogenesis were detected using si-RNA (small interfering RNA) transfection. In results, Tiam1 protein showed a cytoplasmic and nuclear staining pattern in cervical cancer tissues. The strongly positive expression of Tiam1 protein was observed in 51.72% (90/174) of cervical cancers, which was significantly higher than in CINs and normal cervical epithelia tissues (9.38%, 3/32). High Tiam1 protein expression was closely associated with advanced clinical stage, differentiation, lymph node (LN) metastasis, HPV infection and lower overall survival (OS) rates in cervical cancer. Multivariate analysis indicated that Tiam1 was an independent prognostic factor, along with clinical stage, in patients with cervical cancer. Additionally, Tiam1 depletion by RNAi in cervical cancer cells significantly decreased cell proliferation, migration and angiogenesis. In conclusion, our study demonstrated that upregulation of Tiam1 contributes to cervical cancer disease progression and indicates poor survival outcome.

HBXIP suppression reduces cell proliferation and migration and its overexpression predicts poor prognosis in non-small-cell lung cancer.

Emerging evidence has demonstrated that the high expression of HBXIP has been correlated with many cancers. With evaluation of the functional role of HBXIP in non-small-cell lung cancer, the primary aim of this study is to investigate the correlation between HBXIP expression and the prognosis of non-small-cell lung cancer patients. The protein levels of HBXIP were detected using western blotting in non-small-cell lung cancer cells. Cell proliferation and migration assays were measured to evaluate the function of HBXIP in non-small-cell lung cancer cells. A total of 120 non-small-cell lung cancer patients with strict follow-up and 60 adjacent non-tumor lung tissues were selected for immunohistochemical staining of the HBXIP protein. The localization of the HBXIP protein was detected in A549 non-small-cell lung cancer cells using immunofluorescence staining. The correlation between HBXIP expression and the clinicopathological features of non-small-cell lung cancer patients was analyzed by a chi-squared and Fisher's exact test. The overall survival rates of all of the non-small-cell lung cancer patients were calculated using the Kaplan-Meier method, and univariate and multivariate analyses were performed using the Cox proportional hazards regression model. In function, we showed that suppression of HBXIP decreased A549 cell proliferation and migration. HBXIP protein showed a mainly cytoplasmic staining pattern in non-small-cell lung cancer using immunohistochemical staining in paraffin-embedded non-small-cell lung cancer tissues and immunofluorescence staining in A549 cells. The HBXIP protein had strong positive staining in the non-small-cell lung cancer tissues, which was significantly higher than the percentage of adjacent non-tumor tissues. The overexpression of HBXIP was closely correlated with histological grade, clinical stage, lymph node metastasis, and lower overall survival rates of patients with non-small-cell lung cancer. Moreover, multivariate analysis suggested that HBXIP emerged as a significant independent prognostic factor along with clinical stage in patients with non-small-cell lung cancer. In conclusion, a high level of expression of HBXIP is associated with the progression of non-small-cell lung cancer and may be a useful biomarker for poor prognostic evaluation and a potential molecular therapy target for patients with non-small-cell lung cancer.

HBXIP overexpression is correlated with the clinical features and survival outcome of ovarian cancer.

BACKGROUND: Accumulated evidence has demonstrated that Mammalian hepatitis B X-interacting protein (HBXIP) has broad roles in cancer. Although HBXIP is associated with a variety of cancers, the HBXIP protein expression level and its clinical significance in ovarian cancer have not yet been determined. The aim of this study is to investigate the association between HBXIP expression and the clinicopathological features of ovarian cancer patients to determine whether HBXIP may be correlated with a poor prognosis in ovarian cancer patients. METHODS: HBXIP protein expression was assessed in a well-characterized series of ovarian cancer tissue samples (n = 120) with long-term follow-up, using immunohistochemistry to determine the location pattern and expression of HBXIP in ovarian cancer. The localization of HBXIP was detected in SKOV-3 ovarian cancer cells using immunofluorescence (IF) staining. The relationship between high HBXIP expression and the clinicopathological features of ovarian cancer patients was analyzed by Chi-square and Fisher's exact test. Overall survival (OS) rates of all the ovarian cancer patients were calculated using the Kaplan-Meier method, and univariate and multivariate analyses were performed using the Cox proportional hazards regression model. RESULTS: IF staining revealed strongly positive signals for HBXIP in both cytoplasm and nucleus, but mainly in the cytoplasm of SKOV-3 ovarian cancer cells. High HBXIP expression was predominantly observed in ovarian cancer tissues but not the adjacent non-tumor ovarian tissues. The strongly positive rate of HBXIP expression was 60.0% (72/120) in ovarian cancer and was significantly higher than in adjacent non-tumor tissues (17.4%, 4/23) (P = 0.000). High HBXIP expression was positively correlated with the occurrence of lymph node metastases (P = 0.025), histological grade (P = 0.036) and clinical stage (P = 0.003). The patients with high HBXIP expression had lower overall survival (OS) rates. Moreover, multivariate analysis indicated that HBXIP, in addition to the clinical stage, was a significant independent prognostic factor in patients with ovarian cancer. CONCLUSIONS: High-level expression of HBXIP is associated with the progression of ovarian cancer and may be an effective biomarker for poor prognostic evaluation as well as a potential molecular therapy target for ovarian cancer patients.

Mortalin overexpression predicts poor prognosis in early stage of non-small cell lung cancer.

Mortalin is a member of the heat shock protein 70 family, which is involved in multiple cellular processes and may play key roles in promoting carcinogenesis. This study attempted to identify the clinical consequences of Mortalin overexpression and its roles in the prognostic evaluation of non-small cell lung cancer. A total of 120 non-small cell lung cancer samples paired with the adjacent non-tumor tissue samples and 10 normal lung tissues were selected for immunohistochemical staining for Mortalin. The localization of Mortalin was detected in A549 non-small cell lung cancer cells using immunofluorescence staining. The correlations between Mortalin overexpression and the clinical features of non-small cell lung cancers were evaluated using the chi-square test. The survival analysis was calculated via the Kaplan-Meier method and the Cox proportional hazard models. Our studies suggested that Mortalin exhibited a primarily cytoplasmic staining pattern in the non-small cell lung cancers. The rate of strongly positive Mortalin expression was higher in the non-small cell lung cancer samples than in the adjacent non-tumor samples or in normal lung tissues. Mortalin overexpression was significantly correlated with high histological grades, advanced stages, lymph node metastases, and lower disease-free survival and overall survival rates of the patients with non-small cell lung cancer. The survival analysis demonstrated that Mortalin overexpression was a significant independent prognostic factor in non-small cell lung cancer, especially for patients with early stage of non-small cell lung cancer. In conclusion, Mortalin is up-regulated in non-small cell lung cancer, and it may be a potential biomarker of prognostic evaluation and a molecular therapeutic target for patients with early stage of non-small cell lung cancer.

HBXIP over expression as an independent biomarker for cervical cancer.

BACKGROUND: Emerging evidence demonstrated that hepatitis B virus X-interacting protein (HBXIP) has broad roles in cancers. However, high-level expression of HBXIP has been correlated with human malignancies, suggesting roles in carcinogenesis and tumor progression. The aim of the study is to investigate the role and mechanism of HBXIP oncogene and the correlation to the clinicopathological status in cervical cancers. METHODS: A total of 107 cervical cancer patients with strict follow-up, 105 cervical intraepithelial neoplasia (CIN) and 31 normal cervical epithelia samples were selected for immunohistochemical (IHC) staining of HBXIP protein. Additionally, the cervical cancer cell line of SiHa was included in this study. The relationship between HBXIP expression and clinicopathological characteristics were analyzed to verify the clinical value of HBXIP protein expression in patient prognosis, and survival rates were calculated using the Kaplan-Meier method. RESULTS: HBXIP protein showed a mainly cytoplasmic staining pattern in cervical cancers by using IHC staining in paraffin embedded cervical cancer tissues and IF staining in SiHa cervical cancer cells. The strongly positive rate of HBXIP protein expression was significantly higher in cervical SCCs and CINs than in normal cervical epithelia. HBXIP protein over-expression was significantly correlated with the clinical stage, differentiation, lymph node metastasis, HPV infection, the over-expression of P63 and overall survival rates in cervical cancer. All of these data defined that HBXIP was involved in the progression of the cervical cancer. However, the detailed mechanism need to the further study. CONCLUSIONS: HBXIP over-expression appears to associate with cervical cancer progression, and may potentially be used as a cervical cancer biomarker for the early diagnosis, prognostic evaluation and therapeutic target for cervical cancer.

The clinicopathological significance of Mortalin overexpression in invasive ductal carcinoma of breast.

BACKGROUND: Mortalin/GRP75 is a ubiquitous mitochondrial chaperone which related to the cytosolic heat shock protein 70 (HSP70), and plays a role in carcinogenesis. This study aims to investigate the Mortalin expression in breast cancer and its correlation with the outcome of the patients with breast cancer. METHODS: A total of 155 invasive ductal carcinoma of breast patients with strict follow-up, 52 ductal carcinoma in situ (DCIS) and 45 adjacent non-tumor breast tissues were selected for immunohistochemical (IHC) staining of Mortalin protein. The localization of Mortalin protein was detected in MDA-MB231 breast cancer cells using immunofluorescence (IF) staining. The correlations between overexpression of Mortalin and the clinical features of patients with breast cancer were evaluated using chi-square test and Fisher's exact tests. The survival rates were calculated by the Kaplan-Meier method, and the relationship between prognostic factors and patient survival was also analyzed by the Cox proportional hazard models. RESULTS: Mortalin protein showed a mainly cytoplasmic staining pattern in breast cancers by using IHC staining in paraffin embedded breast cancer tissues and IF staining in MDA-MB231 breast cancer cells. The strongly positive rate of Mortalin protein was 63.9% (99/155) in invasive ductal carcinoma of breast and was significantly higher than in DCIS 34.6% (18/52) and adjacent non-tumor tissues 15.6% (7/45). Overexpression of Mortalin was closely correlated with histological grade, clinical stage, lymph node metastasis, lower disease free survival (DFS) and overall survival (OS) rates of patients with breast cancer. Moreover, multivariate analysis suggested that Mortalin emerged as a significant independent prognostic factor along with clinical stage and Her2 expression status in patients with breast cancer. CONCLUSIONS: Mortalin is upregulated in breast cancer, and may be a useful poor prognostic biomarker as well as a potential therapeutic target for patients with breast cancer.