PTEN Deficiency Induces an Extrahepatic Cholangitis-Cholangiocarcinoma Continuum via Aurora kinase A in Mice.

BACKGROUND & AIMS: The PTEN-AKT pathway is frequently altered in extrahepatic cholangiocarcinoma (eCCA). We aim to evaluate the role of PTEN in the pathogenesis of eCCA and find novel therapies for this disease. METHODS: The Pten gene in the biliary epithelial cells were genetically deleted using the Cre-loxp system. The pathologies were evaluated both macroscopically and histologically. The characteristics were further analyzed by immunohistochemistry (IHC), RT-PCR, cell culture, and RNAseq. Some features were compared to those in human eCCA samples. Further mechanistic studies utilized the conditional knockout of Trp53 and Aurora kinase A (Aurka) genes. Experimental therapy was tested using an Aurka inhibitor. RESULTS: We observed that genetic deletion of the Pten gene in the extrahepatic biliary epithelium and peri-ductal glands initiated sclerosing cholangitis-like lesions in mice, resulting in enlarged and distorted extrahepatic bile ducts in mice as early as one month old. Histologically, these lesions exhibited increased epithelial proliferation, inflammatory cell infiltration, and fibrosis. With aging, the lesions progressed from low-grade dysplasia to invasive carcinoma. Trp53 inactivation further accelerated the disease progression, potentially through downregulating senescence. Further mechanistic studies showed that both human and mouse eCCA showed high expressions of AURKA. Notably, the genetic deletion of Aurka completely eliminated Pten deficiency-induced extrahepatic bile duct lesions. Furthermore, pharmacological inhibition of Aurka alleviated disease progression. CONCLUSIONS: Pten deficiency in extrahepatic cholangiocytes and peribiliary glands led to a cholangitis-to-cholangiocarcinoma continuum through an Aurka-dependent manner. These findings offer new insights into preventive and therapeutic interventions for extrahepatic CCA. IMPACT AND IMPLICATIONS: The aberrant PTEN-PI3K-AKT signaling pathway is commonly observed in human extrahepatic cholangiocarcinoma (eCCA), a disease with a poor prognosis. In our study, we developed a mouse model mimicking cholangitis to eCCA progression by conditionally deleting the Pten gene via Pdx1-Cre in epithelial cells and peribiliary glands of the extrahepatic biliary duct. The conditional Pten deletion in these cells led to cholangitis, which gradually advanced to dysplasia, ultimately resulting in eCCA. The loss of Pten heightened Akt signaling, cell proliferation, inflammation, fibrosis, DNA damage, epigenetic signaling, epithelial-mesenchymal transition (EMT), cell dysplasia, and cellular senescence. Genetic deletion or pharmacological inhibition of Aurka successfully halted the disease progression. This model shall be valuable for testing novel therapies and unraveling the mechanisms of eCCA tumorigenesis.

CLCA1 is Poorly Expressed in Stomach Adenocarcinoma and Correlates with Immune Infiltration / CLCA1 se Expresa Deficientemente en el Adenocarcinoma de Estómago y se Correlaciona con la Infiltración Inmune

SUMMARY: Calcium-activated chloride channel regulator 1 (CLCA1) is associated with cancer progression. The expression and immunologic function of CLCA1 in stomach adenocarcinoma (STAD) remain unclear. In this investigation, the expression of CLCA1 in STAD tissues and its involvement in the progression and immune response of STAD were examined using databases such as cBioPortal, TISIDB, and UALCAN. In order to validate the expression level of CLCA1 protein in gastric adenocarcinoma, thirty clinical tissue specimens were gathered for immunohistochemical staining. The findings indicated a downregulation of CLCA1 in STAD patients, which was correlated with race, age, cancer grade, Helicobacter pylori infection, and molecular subtype. Through the examination of survival analysis, it was identified that diminished levels of CLCA1 within gastric cancer cases were linked to decreased periods of post-progression survival (PPS), overall survival (OS), and first progression (FP) (P<0.05). The CLCA1 mutation rate was lower in STAD, but the survival rate was higher in the variant group. The correlation between the expression level of CLCA1 and the levels of immune infiltrating cells in STAD, as well as the immune activating molecules, immunosuppressive molecules, MHC molecules, chemokines, and their receptor molecules, was observed. Gene enrichment analysis revealed that CLCA1 may be involved in STAD progression through systemic lupus erythematosus (SLE), proteasome, cell cycle, pancreatic secretion, and PPAR signaling pathways. In summary, CLCA1 is anticipated to function as a prognostic marker for patients with STAD and is linked to the immunization of STAD.

El regulador 1 del canal de cloruro activado por calcio (CLCA1) está asociado con la progresión del cáncer. La expresión y la función inmunológica de CLCA1 en el adenocarcinoma de estómago (STAD) aún no están claras. En esta investigación, se examinó la expresión de CLCA1 en tejidos STAD y su participación en la progresión y respuesta inmune de STAD utilizando bases de datos como cBioPortal, TISIDB y UALCAN. Para validar el nivel de expresión de la proteína CLCA1 en el adenocarcinoma gástrico, se recolectaron treinta muestras de tejido clínico para tinción inmunohistoquímica. Los hallazgos indicaron una regulación negativa de CLCA1 en pacientes con STAD, que se correlacionó con la raza, la edad, el grado del cáncer, la infección por Helicobacter pylori y el subtipo molecular. Mediante el examen del análisis de supervivencia, se identificó que los niveles reducidos de CLCA1 en los casos de cáncer gástrico estaban relacionados con períodos reducidos de supervivencia posterior a la progresión (PPS), supervivencia general (OS) y primera progresión (FP) (P <0,05). La tasa de mutación CLCA1 fue menor en STAD, pero la tasa de supervivencia fue mayor en el grupo variante. Se observó la correlación entre el nivel de expresión de CLCA1 y los niveles de células inmunes infiltrantes en STAD, así como las moléculas activadoras inmunes, moléculas inmunosupresoras, moléculas MHC, quimiocinas y sus moléculas receptoras. El análisis de enriquecimiento genético reveló que CLCA1 puede estar involucrado en la progresión de STAD a través del lupus eritematoso sistémico (LES), el proteasoma, el ciclo celular, la secreción pancreática y las vías de señalización de PPAR. En resumen, se prevé que CLCA1 funcione como un marcador de pronóstico para pacientes con STAD y está vinculado a la inmunización de STAD.

Aberrant expression of UBE2C in endometrial cancer and its correlation to epithelial mesenchymal transition.

Ubiquitin-conjugating enzyme E2C (UBE2C), its overexpression promotes tumor progression, is a key component of the ubiquitin conjugating proteasome complex. Epithelial-mesenchymal transition, which is lost epithelial features and gained mesenchymal features in some epithelial cancers, is involved in epithelial cancers' invasiveness and metastasis. The aim of this study is to detect the expression of UBE2C, WNT5α, and E-cad in endometrial cancer (EC) and their clinical significance. The expression of UBE2C, WNT5α, and ZEB1 in 125 cases EC tissues were detected by immunohistochemistry. Patients clinicopathological, demography, and follow-up data were also collected. Positive rates of expression of UBE2C and ZEB1 were significantly higher in EC tissues when compared with the control tissues. The positive expression of UBE2C and ZEB1 were positively associated with tumor stages, local lymph node metastasis, and International Federation of Gynecology and Obstetrics (FIGO) stages. The positive rate of expression of WNT5a was significantly lower in EC tissues when compared with the control tissues. And positive expression of E-cad was inversely related to tumor stages, lymph node metastasis stages, and FIGO stages. Kaplan-Meier analyses demonstrated that positive expression of UBE2C or ZEB1 for EC patients had unfavorably overall survival time when compared with patients with negative expression of UBE2C or ZEB1. And EC patients with positive expression of WNT5a had favorably overall survival time when compared with EC patients with negative expression of WNT5a. Multivariate analysis demonstrated that positive expression UBE2C, WNT5α, and ZEB1, as well as FIGO stages were independent prognostic factors for EC patients. UBE2C, ZEB1, and WNT5a should be considered promising biomarkers for EC patients' prognosis.

Menetrier's disease and differential diagnosis: A case report.

BACKGROUND: Gastric mucosal hypertrophy, also known as Menetrier's disease (MD), is more common in men over 50 years of age, and the cause is unknown. The symptoms of the disease are atypical, mostly accompanied by hypoproteinemia and edema, and sometimes accompanied by symptoms such as epigastric pain, weight loss, and diarrhea. Most experts believe that the site of the disease is mainly located in the fundus of the stomach and the body of the stomach. We found that the site of the disease in this patient involved the antrum of the stomach. CASE SUMMARY: We introduced the case of a 24-year-old woman who had repeated vomiting for 5 d and was admitted to our hospital. After various examinations such as computed tomography and pathology in our hospital, the final diagnosis of the presented case is MD. The salient feature is that the mucosal folds in the fundus and body of the stomach are huge and present in the shape of gyrus. The greater curvature is more prominent, and there are multiple erosions or ulcers on the folds. The patient did not undergo gastric surgery and did not undergo re-examination. She is drinking Chinese medicine for treatment, and her vomiting and abdominal pain symptoms have improved. This disease is relatively rare in clinical practice, and it is easy to be misdiagnosed as gastric cancer, chronic gastritis and gastric lymphoma, etc. CONCLUSION: MD can occur in the antrum, it is necessary to raise awareness of the disease and reduce misdiagnosis.

Apatinib combined with concurrent chemoradiotherapy in patients with subglottic small cell carcinoma: a case report.

Subglottic small cell carcinoma (SSMCC) is a rare type of neoplasm, meaning that laryngeal cancer guidelines in several countries, including the National Comprehensive Cancer Network (NCCN) guidelines, do not include treatment principles for SSMCC. Angiogenesis is an established factor in tumor initiation, growth, and dissemination. Apatinib mesylate, an orally administered drug, is a novel inhibitor of vascular endothelial growth factor receptor-2, a key mediator of angiogenesis, and has been shown to be safe and efficacious in the treatment of certain types of malignant tumors. To the best of our knowledge, there are few reports on the treatment of SSMCC with apatinib combined with concurrent chemoradiotherapy. In the present report of SSMCC in a 64-year-old woman, oral apatinib was given daily at a dose of 250 mg in combination with concurrent chemoradiotherapy; the only toxicities reported were mild leukopenia and finger numbness. Clear and rapid efficacy was observed with the disappearance of the tumor mass. Our findings indicate that apatinib combined with concurrent chemoradiotherapy may be effective in patients with SSMCC, with adverse reactions within the manageable range, thus representing an additional treatment option for this type of malignancy.

Expression of Formin-like 2 and cortactin in gallbladder adenocarcinoma and their clinical significance.

OBJECTIVE: To investigate the expressions of Formin-like 2 (FMNL2) and Cortactin (CTTN) in gallbladder adenocarcinoma (GBAC) and their associations with the clinicopathological characteristics of the patients. METHODS: The expressions of FMNL2 and CTTN were detected with immunohistochemistry (Max Vision) in 105 GBAC tissues and 40 normal gallbladder tissues. RESULTS: The positive expression rates of FMNL2 and CTTN in normal gallbladder tissues were 25% and 20%, different from the positive expression rates of 84.76% and 86.67% in GBAC tissues (P < 0.001). The positive expression rate of FMNL2 and CTTN in GBAC correlated with tumor differentiation, tumor-node-metastasis (TNM), lymph node metastasis (LNM), and distant metastasis. FMNL2 expression was positively correlated with CTTN expression. Kaplan-Meier analysis showed that the overall survival time of patients with positive expressions group of FMNL2 and CTTN was significantly shorter than that of the negative expression group. Cox multivariate analysis showed that TNM, LNM, distant metastasis, and positive expression of FMNL2 and CTTN were independent factors influencing the prognosis of patients with GBAC (P < 0.05). CONCLUSION: The positive expression of FMNL2 and CTTN in GBAC is significantly increased, which may be related to the occurrence and development of GBAC. The combined detection of FMNL2 and CTTN may provide a scientific theoretical basis for the early diagnosis of GBAC, the development of new antitumor drugs, and the search for new targets of biotherapy.

FOXM 1 induces Vasculogenic mimicry in esophageal cancer through ß-catenin /Tcf4 signaling.

OBJECTIVE: To investigate the role of FOXM1, ß-catenin and TCF4 in esophageal cancer (EC) and their relationship to VM (Vasculogenic Mimicry). METHODS: CCK-8 were performed to examine EC cell proliferation in FOXM1 silenced cells. EC cell migration and invasion were investigated through wound healing and Transwell assays, respectively. The formation of pipe like structures were assessed in 3D cultures. The expression of Foxm1, ß-catenin, Tcf4 and E-cadherin were investigated through western blot, RT-qPCR and immunohistochemistry (IHC) staining. The relationship between FOXM1 expression, clinic-pathological features, and overall survival (OS) were further analyzed. RESULTS: A loss of FOXM1 expression correlated with the OS of ESCC patients. FOXM1 silencing led to a loss of cell growth and suppressed cell migration and invasion in ESCC cells. VM structures were identified in ESCC tissues and human EC cell lines. Mechanistically, FOXM1 was found to promote tumorigenesis through the regulation of ß-catenin, Tcf4, and E-cadherin in EC cells, leading to the formation of VM structures. CONCLUSIONS: These findings highlight FoxM1 as a novel therapeutic target in ESCC.

Inhibition of KiSS-1 on metastasis of nasopharyngeal carcinoma implant tumor in nude mice.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a kind of head-neck malignant neoplasm originated from the nasopharyngeal epithelium and is mainly prevalent in Southern China and Southeast Asia countries. KiSS-1 is an inhibitor of tumor metastasis in a range of cancers. METHODS: We establish a cell substrain of SUNE-1-5-8F (NPC cell line from humans) that trsnfected with lentiviral vectors carried with KiSS-1 gene and were selected by puromycin. A transplantation tumor animal model in BALB/c-nu mice was successfully established with a substrain that stably overexpressed KiSS-1. RESULTS: Our result showed that the size of transplantation tumor in the nude mice with KiSS-1 overexpression in transplantation tumor was not difference from the size of transplantation tumor in the controlled transplantation tumor mice. We detected metastatic tumor in lung but not in liver. Moreover, we also found that in the nude mice with KiSS-1 overexpression in transplantation tumor showed extremely fewer metastatic tumor in lung compared with the controlled transplantation tumor mice model. In conclusion, KiSS-1 may be beneficial for the inhibition of metastasis of human NPC. CONCLUSION: This study may throw light on the treatment of NPC and may help improve the prognosis of patients with NPC.

Evaluation of the correlativity of gender determining region Y-box 4, N-cadherin, CD44 and E-cadherin expression in the prognosis of esophageal squamous cell carcinoma.

BACKGROUND: SOX4 is highly expressed in many different tumor types, and SOX4 has been reported in the literature to participate in tumor proliferation, damaging and movement by leading Epithelial-Mesenchymal Transition. Cancer vital cells and Epithelial-Mesenchymal Transition have been repeatedly confirmed to participate during the proliferation, damaging and movement of cancer. This research examined the association of the Epithelial-Mesenchymal Transition-related molecules E-cadherin, N-cadherin, CD44, and SOX4 in the ESCC and aimed for providing inspiration for clinical treatment as well as to indicate a new direction for detecting invasion and forecasting the prospect of affected role using ESCC. METHODS: Immunohistochemistry was utilized to observe the expression of the S0X4, N-cadherin, CD44 and E-cadherin proteins. Survival analysis of the positive and negative SOX4, E-cadherin, N-cadherin and CD44 protein expression groups was performed by the Kaplan-Meier approach. OUTCOMES: A confirming relationship was observed among the expression of SOX4, N-cadherin or CD44 and tumor diameter, distant metastasis, deepness of damaging, lymph node metastasis, pTNM stage and histological grade (P<0.05). Spearman correlativity calculation displayed that the expression of the SOX4 protein was obviously responded with the expression of the N-cadherin and CD44 proteins. Moreover, the expression of the N-cadherin and CD44 proteins was also positively correlated. The E-cadherin protein was negatively correlated with SOX4, N-cadherin and CD44 protein expression in ESCC. SOX4, N-cadherin, CD44, E-cadherin, age and distant metastasis were determined to be separate elements that influenced the prognosis of patients with ESCC. CONCLUSIONS: We found that suppression of ESCC providers can suppress the growth of bad tumors and change therapeutic results for ESCC patient since CD44 supports the induction of Epithelial-Mesenchymal Transition in ESCC.

Aberrant expression of MYH9 and E-cadherin in esophageal squamous cell carcinoma and their relationship to vasculogenic mimicry.

PURPOSE: To investigate whether vasculogenic mimicry (VM) exists in esophageal squamous cell carcinoma (ESCC) and to elucidate the relationship among expression of MYH9, E-cadherin and VM. METHODS: The expression of MYH9 (non-muscle myosin heavy chain 9), E-cadherin protein and VM in 120 specimens of esophageal squamous cell carcinoma (ESCC) and 120 specimens of normal esophageal mucosa were detected by using immunohistochemical and histochemical staining. RESULTS: VM channels were identified in 58 (48.33%) of the 120 ESCC specimens and none of the normal esophageal mucosa was found to have VM. The rates of expression of MYH9 and E-cad in ESCC were 57.50% and 40.00%, while rates in the control group were 13.33% and 73.33%, respectively (P<0.05). VM and the expression levels of MYH9 and E-cad were significantly connected with lymph node metastasis, serosa invasion, pTNM staging and 5-year-survival rates of patients with ESCC (P<0.05). VM was positively correlated with MYH9, but negatively correlated with E-cad, and MYH9 was negatively significantly correlated with E-cad. The 5-year-survival rates of patients with ESCC were 6.89% (4/58) in the VM group and 67.74% (42/62) in the non-VM group, 8.00% (4/50) in high MYH9 expression group and 60.00% (42/70) in low MYH9 expression group. However, the 5-year-survival rate in high E-cad expression group was 86.95% (40/46) and that in low E-cad expression group was 8.11% (6/74) (P<0.05). Cox multifactorial regression analysis demonstrated that lymph node metastasis, pTNM stage, VM and expression levels of MYH9 and E-cad were independent risk factors in patients with ESCC (P<0.05). CONCLUSION: ESCC'patients with VM had a poor differentiation and a bad clinical prognosis; Combined detection of VM, MYH9 and E-cad may play an essential role in predicting the invasion, metastasis, and progression of patients with ESCC.

[Expressions of ΔNp63α, DPC4/Smad4 and P21 in cervical squamous cell carcinoma an their clinical significance].

OBJECTIVE: To investigate the expressions of ΔNp63α, DPC4/Smad4 and P21 in cervical squamous cell carcinomas and explore their implications in tumorigenesis, progression and prognosis of the malignancy. METHODS: The expressions of ΔNp63α, DPC4/Smad4 and P21 were examined with immunohistochemistry in 100 specimens of cervical squamous cell carcinoma, 40 specimens of cervical intraepithelial neoplasia (CIN) and 40 specimens of normal cervical tissues to explore their associations with the occurrence, progression and prognosis of cervical squamous cell carcinoma. RESULTS: The expressions of ΔNp63α and DPC4/Smad4 decreased and P21 expression increased significantly in the order of normal cervical tissue, CIN and cervical squamous cell carcinoma (P < 0.01), and their expressions were associated with the differentiation, clinical stages and lymph node metastasis of cervical squamous cell carcinoma (P < 0.01). The expression of ΔNp63α was positively correlated with the expression of DPC4/Smad4 (r=0.581, P < 0.05), and they were both negatively correlated with P21 expression (r=-0.449 and -0.254, respectively; P < 0.05). Kaplan-Meier survival analysis showed that patients with cervical squamous cell carcinoma positive for ΔNp63α and DPC4/Smad4 had a significantly higher 5-year survival rate than those negative for ΔNp63α and DPC4/Smad4 (P < 0.001); the patients positive for P21 had a significantly lower 5-year survival rate than the P21-negative patients (P < 0.005). CONCLUSIONS: The expressions of ΔNp63α, DPC4/Smad4 and P21are related with the differentiation, invasion, lymph node metastasis, pTNM stage and prognosis of in cervical squamous cell carcinomas, suggesting their value as potential markers for prognostic evaluation of patients with cervical squamous cell carcinoma.

Associations of MACC1, AGR2, and KAI1 expression with the metastasis and prognosis in head and neck squamous cell carcinoma.

BACKGROUND: Metastasis-associated in colon cancer-1 (MACC1, was firstly found in colon cancer and associated metastasis and prognosis in various cancers), anterior gradient 2 (AGR2, was considered as a valuable prognostic factor for some cancers), and Kangai 1 (KAI1, was a tumor metastasis suppressor gene) are all related to metastasis and prognosis of many cancers. However, the associations of MACC1, AGR2, and KAI1 in head and neck squamous cell carcinoma (HNSCC) are still unclear. In this study, we analyzed associations among MACC1, AGR2, and KAI1 in HNSCC, and their respective associations with clinicopathological parameters and overall survival (OS) in HNSCC. METHODS: Positive expression of MACC1, AGR2, and KAI1 in 106 whole HNSCC tissue samples was detected by immunohistochemical staining. Patient's clinical data and demographics were both collected. RESULTS: Positive rates of MACC1 and AGR2 were significantly higher, and positive rate of KAI1 was significantly lower, in HNSCC and than those in control tissues. Positive rates of MACC1 and AGR2 were positively correlated with grades of tumor, TNM stages, and lymph node metastasis (LNM) stages, and negatively with patients OS; positive rate of KAI1 was negatively associated with grades of tumor, TNM stages, and LNM stages, and the positive expression of KAI1 subgroup had significantly longer OS than did the negative KAI1 subgroup. In multivariate analysis, positive expression MACC1, AGR2, and KAI1, and tumor stages, as well as LNM stages were potential to be independent prognostic factors for OS in patients with HNSCC. CONCLUSIONS: MACC1, AGR2, and KAI1 may represent potential metastatic and prognostic biomarkers, as well as promising therapeutic targets for HNSCC.

Value of CagA, HER2, ALDH1, and KiSS-1 in predicting metastasis and prognosis for gastric adenocarcinoma.

BACKGROUND: Cytotoxin-associated gene A (CagA), is able to translocate into gastric epithelial cells. Human epidermal growth factor receptor 2 (also named as HER2, is a proto-oncogene which can encode a transmembrane receptor), Aldehyde dehydrogenase 1 (ALDH1, a biomarker of cancer stem cells), and KiSS-1 (a suppressor gene of cancer metastasis) are all valuably predictive biomarkers for various human cancers. The purpose of this study is to investigate the associations among CagA, HER2, ALDH1, and KiSS-1 in gastric adenocarcinoma (GAC), and their respective associations with clinical characteristics and survival in GAC. METHODS: The expression of CagA, HER2, ALDH1, and KiSS-1 in 232 cases of whole GAC tissues were detected by immunohistochemical staining. Patient clinical and survival data were also collected. RESULTS: Positive expression of CagA, HER2, and ALDH1 is significantly higher, and positive expression of KiSS-1 is significantly lower, in GAC tissues than in the corresponding normal tissues. Furthermore, the positive expression of CagA, HER2, ALDH1, and KiSS-1 were significantly associated with tumor grade, tumor stage, lymph node metastasis (LNM) stage, and tumor node metastasis (TNM) stages, and with patients' overall survival (OS); whereas the KiSS-1 positive group had longer OS than did the KiSS-1 negative group. In logistic analysis, positive expression of CagA, HER2, ALDH1, and KiSS-1 are significantly associated with LNM of patients with GAC. COX regression analysis indicated that positive expression of CagA, HER2, ALDH1, and KiSS-1, and tumor stages, LNM stages, and TNM stages were independent prognostic factors for patients with GAC. CONCLUSIONS: Expression of CagA, HER2, ALDH1, and KiSS-1 should be considered as promising biomarkers for metastasis and prognosis, as well as potential therapeutic targets for GAC.

The expressions of CD133, ALDH1, and vasculogenic mimicry in osteosarcoma and their clinical significance.

BACKGROUND AND PURPOSE: Osteosarcoma is an aggressive malignant bone tumor in children and adolescents, which is more likely to recur and metastasize at the early stages. Cancer stem cells (CSC, CD133 is a biomarker of cancer stem cells), angiogenesis, and vasculogenic mimicry (VM) are closely related to tumor metastasis and recurrence. In this study, we investigated the associations among CD133, aldehyde dehydrogenase 1 (ALDH1), and VM in osteosarcoma, and their associations with clinical characteristics. METHODS: Positive rates of CD133, ALDH1, and VM in 96 whole osteosarcoma tissue samples were detected by immunohistochemistry (IHC) and histochemistry staining. Patients' clinical data were also collected. RESULTS: Positive rates of CD133, ALDH1, and VM were significantly higher in osteosarcoma tissues compared with the control tissues. Positive rates of CD133, ALDH1, and VM were positively associated with lymph node metastasis, distant metastasis, Enneking stages, and patients' overall survival (OS). A multivariate analysis indicated that the positive rates of CD133, ALDH1, and VM, as well as the Enneking stages were independent prognostic factors of osteosarcoma. CONCLUSION: The positive rates of CD133, ALDH1, and VM could represent potential biomarkers for metastasis and prognosis, which suggests these molecules might be promising therapeutic targets for osteosarcoma.

Over-expression of Thrombospondin 4 correlates with loss of miR-142 and contributes to migration and vascular invasion of advanced hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a common malignancy found worldwide and is associated with a high incidence of metastasis and vascular invasion. Elucidating the molecular mechanisms that underlie HCC tumorigenesis and progression is necessary for the development of novel therapeutics. By analyzing the Cancer Genome Atlas Network (TCGA) dataset, we identified Thrombospondin 4 (THBS4) is significantly overexpressed in HCC samples and is correlated with prognosis. Overexpression of THBS4 was also highly correlated with vascular invasion of advanced HCC. While THBS4 is often overexpressed in HCC it has also been shown to inhibit tumor growth by mediating cell-to-cell and cell-to-matrix interactions. Here, we identified that knockdown of THBS4 inhibits migration and invasion of HCC cells and inhibits HCC induced angiogenesis. MiRNAs are crucial regulators of multiple cellular processes, and aberrant expression of miRNAs has been observed to effect cancer development and progression. We further found that miR-142 is an upstream regulator of THBS4 in HCC cells. Moreover, miR-142 was significantly down-regulated in HCC tissue samples and correlated with overexpression of THBS4. Overexpression of miR-142 inhibited invasion and angiogenesis of HCC cells and re-expression of THBS4 overcame these effects of miR-142 expression. Stable over-expression of miR-142 significantly inhibited tumour growth in a xenograft tumour model through inhibiting THBS4 expression and tumor angiogenesis. In conclusion, our findings indicate that loss of miR-142 results in the over-expression of THBS4, which enhances HCC migration and vascular invasion. Thus, targeting THBS4 or miR-142 may provide a promising therapeutic strategy for treatment of advanced HCC.

Expression of LGR5 in oral squamous cell carcinoma and its correlation to vasculogenic mimicry.

BACKGROUND: LGR5, also named as GPR49, is considered as a biomarker of cancer stem cells which have been responsible for the initiation, progression, metastasis, and recurrence of cancers. Vasculogenic mimicry (VM) which defines the formation of fluid-conducting tubes by highly progressive and genetically dysregulated cancer cells has been considered as useful biomarker for metastasis and prognosis in various cancers. In this study, we analyzed associations between LGR5 and VM in oral squamous cell carcinoma (OSCC), and their association with clinicopathological characters in OSCC. METHODS: Positive rates of LGR5 and VM in 190 OSCC tissue samples and correspondence normal tissues were detected by immunohistochemical and histochemical staining. Patients' clinical data were also collected. RESULTS: Positive rates of LGR5 and VM were significantly higher in OSCC tissues than those in normal tissues. Positive rates of LGR5 and VM were positively related to tumor size, grades, lymph node metastasis, and TNM stages, and inversely with patients overall survival time. And there was a positive association between the expression of LGR5 and positive rate of VM. In multivariate analysis, high expression of LGR5 and positive VM and lymph node metastasis, as well as TNM stages were to be considered as independent prognosis factors for overall survival time in patients with OSCC. CONCLUSIONS: The expression of LGR5 and VM represent potential biomarkers for metastasis and prognosis, as well as therapeutic targets for OSCC.

[Expressions of Slug, ZEB1 and KISS-1 in gastric adenocarcinoma and their clinical significance].

OBJECTIVE: To identify potential markers for predicting invasion, metastasis, and prognosis of gastric adenocarcinoma (GAC). METHODS: The expressions of Slug, ZEB1 and KISS-1 were detected immunohistochemically in 261 GAC tissues and 80 normal gastric tissues. RESULTS: The positivity rates of Slug, ZEB1, and KISS-1 in gastric tissues were 2.5%, 1.3%, and 87.5%, respectively, significantly different from the rates of 62.1%, 28.4%, and 41.1% in GAC tissues (P<0.05). The expression level of Slug was significantly correlated with the depth of invasion, lymph node metastasis, and pTNM stages; the positivity rates of both ZEB1 and KISS-1 were significantly correlated with the tumor grade, depth of invasion, lymph node metastasis and pTNM stages. Slug expression was positively correlated with ZEB1 expression, and KISS-1 expression was inversely correlated with Slug and ZEB1 expressions. Kaplan-Meier analysis showed that the overall survival time of patients with positive expressions of Slug and ZEB1 was significantly shorter than that of the negative patients, and the survival time of patients positive for KISS-1 was significantly longer than the negative patients. COX multivariate analysis showed that positive Slug, ZEB1 and KISS-1 protein expressions and pTNM stages were independent prognostic factors of GAC (P<0.05). CONCLUSION: The abnormal expressions of Slug, ZEB1 and KISS-1 may contribute to the tumorigenesis of GAC and are related with lymph node metastasis, pTNM stages, and prognosis of GAC. The combined detection of Slug, ZEB1, and KISS-1 expression has an important value in predicting the progression and prognosis of GAC.

[Expressions of Snail, Slug and KAI1 proteins in cervical carcinoma and their clinicopathological significance].

OBJECTIVE: To explore the expression of Snail and Slug in primary cervical squamous cell carcinoma (CSCC) and their relationship with KAI1 expression. METHODS: The expressions of Snail, Slug, and KAI1 proteins were examined by immunohistochemistry in 154 specimens of CSCC tissues, 50 specimens of cervical intraepithelial neoplasm (CIN), and 40 specimens of normal cervical tissues. RESULTS: The positivity rates of Snail, Slug, and KAI1 expression were 0%, 2.5%, and 95.0% in normal cervical tissues, 32.0%, 34.0% and 64.0% in CIN tissues, and 66.2%, 66.9%, and 43.5% in CSCC tissues, respectively, showing significant differences in the rates among the 3 groups (P<0.05). The expressions of Snail, Slug, and KAI1 were significantly correlated with the histological grades of the tumor, depth of invasion, lymph node metastasis, International Federation of Gynecology and Obstetrics (FIGO) stages, and postoperative survival time (P<0.05). The expressions of Snail and Slug were positively correlated (r=0.752, P<0.001), and both of them were negatively correlated with the expression of KAI1 (P<0.001). Kaplan-Meier analysis showed that patients positive for Snail and Slug had significantly lower survival rates than the negative patients (P<0.001), while a positive expression of KAI1 was associated with a higher survival rate of the patients. Cox regression analysis identified Snail, KAI1, and FIGO stage as independent factors that affected the outcomes of CSCC (P<0.05). CONCLUSION: The expressions of Snail, Slug, and KAI1 are related to the tumor grade, FIGO stage, invasive depth, lymph node metastasis, and prognosis of CSCC, and their combined detection can help estimate the outcomes of the patients.

[T Cell Factor 4, beta-catenin and SFRP1 Expression of Wnt Signaling Pathway in Colorectal Carcinoma and the Prognosis].

Abnormal activation of Wnt signaling pathway is closely related to the occurrence of tumor, and T cell factor 4 (Tcf4 ) and beta-catenin are important signal transmission factors of this pathway. The aim of the present study is to explore the significance and correlation between expression of Tcf4, beta-catenin and secreted frizzled related protein 1(SFRP1), suppressor gene of Wnt signaling pathway, in colorectal carcinoma and their correlations to the clinicopathological factors. The expressions of Tcf4, beta-catenin and SFRP1 were performed with immunohistochemistry staining in 97 cases of primary colorectal carcinoma and 40 cases of normal colorectal mucosa tissues. The results showed that the abnormal expression rates of Tcf4 and beta-catenin in colorectal carcinoma were significantly higher than those in the control groups (P<0.01). The positive rate of SFRP1 was significantly lower than those in the control groups (P<0.01). The abnormal expression rates of Tcf4 and beta-catenin were also related to the lymph node metastasis and Dukes stage (P<0.05). A significant correlation was found between the expressions of SFRP1 and Tcf4, beta-catenin (P<0.05). Overexpression of Tcf4 and beta-catenin was related to poor prognosis (P<0.05). But the survival rates of the group with SFRP1 expressions were higher than those in group without SFRP1 expressions (P<0.05). Cox multifactor regression analysis indicated that Dukes stage, expression of beta-catenin and SFRP1 were independent risk factors of colorectal carcinoma (P<0.05). The results suggested that the abnormal expression of Tcf4 and beta-catenin in colorectal cancer may be related to the reduced or absent expression of SFRP1. beta-catenin accumulation in the nuclei formed complexes with Tcf4 is one of the important molecular switch maintaining colorectal malignant phenotype. The combined detection of these indexes may perform an important role in predicting the progression and prognosis of colorectal cancer, and could provide new molecular targets for gene treatment of colorectal cancer.

[Expression of Tcf-4, MMP7 and Survivin in Colorectal Cancer and Its Clinical Significance].

OBJECTIVE: To explore the clinical and pathological significance and correlations among the xpressions of Tcf-4, MMP7 and survivin in colorectal cancer. METHODS: The expressions of Tcf-4, MMP7 and survivin mRNA in tumor tissues and adjacent normal mucosa from 50 colorectal cancer patients were detected by reverse transcription PCR (RT-PCR). The expressed proteins of Tcf-4, MMP7 and survivin were measured using mmunohistochemistry staining technique (Elivision) in 100 colorectal cancer samples and 60 normal mucosa tissue samples. We analyzed the correlations between those measurements and their associations with clinical and pathological characteristics. RESULTS: Positive expressions of Tcf-4, MMP7 and survivin mRNA were found in both cancer and adjacent mucosa tissues, despite a higher level of expression in the cancer tissues (P < 0.01). Expressed proteins were detected in cancer tissues of 69.00% (69/100) of those with a positive Tcf-4 expression, 77.00% (77/100) of those with a positive MMP7 expression, and 65.00% (65/100) of those with a positive survivin expression. Compared with cancer tissues, lower levels of protein expression were found in normal mucosa tissues [16.67% (10/60) for Tcf-4, 13.33% (8/60) for MMP7 and 15.00% (9/60) for survivin, P < 0.01]. The expressions of Tcf-4, MMP7 and survivin were all associated with lymphatic metastasis and Dukes staging (P < 0.05). MMP7 expression was associated with depth of tumor invasion (P < 0.05). Survivin expression was associated with tumor differentiation. The Spearman rank correlation analyses showed that protein expressions in colorectal cancer tissues in those with a positive Tcf-4 were correlated with those with a positive MMP7 (r = 0.302) and those with a positive survivin (r = 0.279) (P < 0.01), but not in those with a positive MMP7 and those with a positive survivin (r = 0.097, P > 0.05). CONCLUSION: The expression levels of Tcf-4, MMP7 and survivin are high in colorectal cancer, all being linked to lymph node metastasis and Dukes stages of patients. This suggests that they may be involved in the occurrence, development, malignant growth and clinical progression of colorectal cancer.