Overexpression of chloride channel-3 (ClC-3) is associated with human cervical carcinoma development and prognosis.

BACKGROUND: Cervical carcinoma is a major gynecological cancer and causes cancer-related deaths in worldwide, the latent pathogenesis and progress of cervical cancer is still under research. ClC-3 may be an important promoter for aggressive metastasis of malignant tumors. In this research, we explore the ClC-3 expression in cervical carcinoma and its underlying clinical significance, trying to illuminate ClC-3 probable function in the neoplasm malignant behavior, development and prognosis. METHODS: Paraffin-embedded cervical (n = 168) and lymph node (n = 100) tissue specimens were analysed by immunohistochemistry. Fresh human cervical tissue specimens (n = 165) and four human cervical cell lines were tested for ClC-3 mRNA and protein expression levels by quantitative real-time PCR and western blotting. The relationship between the expression levels of ClC-3, the pathological characteristics of the carcinoma, and the clinical prognosis were statistically analysed. RESULTS: In normal and precancerous (LSIL, HSIL) cervical tissues as well as cervical carcinoma tissues, both ClC-3 mRNA and protein expression levels increased significantly (p < 0.05). The expression level of ClC-3 was closely-related to the histological differentiation (p = 0.029), tumour staging (p = 0.016), tumour size (p = 0.039), vascular invasion (p = 0.045), interstitial infiltration depth (p = 0.012), lymphatic metastasis (p = 0.036), and HPV infection (p = 0.022). In an in vitro experiment, ClC-3 mRNA and protein were found to be overexpressed both in the HeLa and SiHa cell lines, but low expression levels were detected in the C-33A and H8 cell lines (p < 0.05). Furthermore, the high expression levels of ClC-3 was significantly correlated to poor survival in cervical carcinoma patients (Log-rank test, p = 0.046). CONCLUSIONS: These data suggest that overexpression of ClC-3 is closely associated with human cervical carcinoma progression and poor prognosis; this suggests that ClC-3 may function as a patent tumour biomarker and a latent therapeutic target for cervical carcinoma patients.

Tumor-associated macrophages correlate with progesterone receptor loss in endometrial endometrioid adenocarcinoma.

AIM: It has been well established that tumor-associated macrophages (TAMs) play a tumor promoting role in endometrial endometrioid adenocarcinoma (EEC). But the association with TAMs and sex hormone receptor expression, and progression of precancerous endometrial lesions in EEC has been little reported. MATERIAL AND METHODS: We used immunohistochemistry to examine the expression of CD68, CD34, vascular endothelial growth factor (VEGF), estrogen receptor (ER) and progesterone receptor (PR) in 95 cases of EEC, as well as 35 cases of endometrial hyperplasia (including 15 atypical hyperplasia, 10 complex hyperplasia and 10 simple hyperplasia). We also correlated TAMs count with various clinicopathological factors, sex hormone receptor, and prognostic value in patients with EEC. RESULTS: We identified that TAMs count increased linearly with disease progression (mean count per case at × 200 magnification: simple hyperplasia, 6.30; complex hyperplasia, 11.20; atypical hyperplasia, 29.40; EEC 55.81, respectively; P < 0.001), that microvascular density (MVD) also increased accordingly (27.50, 30.20, 50.13 and 59.94, respectively; P < 0.001). The expression of progesterone receptor, not of estrogen receptor, significantly decreased with disease progression (P < 0.05). Moreover, histopathologic grades, International Federation of Gynecology and Obstetrics (FIGO) stage (2009), depth of myometrial invasion, pelvic lymph node metastasis, lymphovascular space invasion, and expression of PR and VEGF were associated with TAMs count (P = 0.0001, P = 0.004, P = 0.0001, P = 0.04, P = 0.0001, P = 0.0001, P = 0.0001, respectively). Progesterone receptor expression was also associated with histopathologic grades, lymphovascular space invasion, VEGF and high TAMs (P = 0.035, P = 0.022, P = 0.014, P = 0.001, respectively). The estimated 5-year survival rate of patients with low TAMs was significantly higher than those with high TAMs (96.4% vs 69.8%, P = 0.002). CONCLUSION: TAMs are potentially related to PR loss and progression of precancerous endometrial lesions in EEC.

Tumor-associated macrophages, epidermal growth factor receptor correlated with the triple negative phenotype in endometrial endometrioid adenocarcinoma.

It has been well established that tumor-associated macrophages (TAMs) play a tumor-promoting role in endometrial endometrioid adenocarcinoma (EEC). However, the association with TAMs and the triple-negative phenotype (TNP) in EEC has not yet been reported. We used immunohistochemistry to examine the expression of CD68, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and epidermal growth factor receptor (EGFR) in 186 cases of EEC. Fluorescent in situ hybridization (FISH) was also used for HER2 amplification, and the association with TAMs count, EGFR expression, and triple-negative phenotype was analyzed. Twenty-eight of 186 patients (15.05%) had the TNP. It was associated with advanced stage disease (P<0.0001), high grade disease (P<0.0001), depth of myometrial invasion (P=0.003), pelvic lymph node metastasis (P<0.001), lymphovascular space invasion (P=0.001), and EGFR expression (P=0.032). Margin TAMs count was also significantly increased in the TNP-positive group, the EGFR-positive group, and the PR-negative group (P<0.001, respectively). The TNP was associated with a significantly worse overall survival (OS) (log rank test, P=0.018). The estimated 5-year OS of patients with TNP was 59.1%, while that without TNP was 78.5%. Multivariate analysis showed high margin TAMs, and the histopathological grades were significantly associated with OS. The TNP in EEC is associated with poor prognostic surgical-pathological factors, worse prognosis, as well as with high margin TAMs and overexpression of EGFR, which may serve as potential targeted therapies for the special phenotype in EEC.

[Expression of ICAM-1 (CD54) in pediatric tumor and acute leukemia and its clinic significance in immunotherapy with CIK cell].

This study was aimed to investigate the expression of ICAM-1 (CD54) in pediatric tumor and acute leukemia (AL), so as to understand the distribution of ICAM-1 and its clinical significance. The expression of ICAM-1 in tissues of 46 pediatric tumor patients were detected by immunohistochemistry, and in bone marrow cells of 60 pediatric acute leukemia (AL) patients were detected by flow cytometry. 46 pediatric tumor patients included 10 lymphoma, 3 hepatoblastoma, 6 neuroblastoma, 2 rhabdomyosarcoma, 6 Ewing's bone sarcoma, 2 fibrosarcoma, 5 primitive neuroectodermal tumor, 11 nephroblastoma and 1 osteosarcoma. 60 AL pediatric patients included 20 acute lymphocytic leukemia (ALL) patients and 40 acute nonlymphocytic leukemia (ANLL) patients containing 20 M1, M2, M3 patients and 20 M4, M5. The results indicated that expression of ICAM-1 was more positive in all 3 hepatoblastoma cases, which represent a higher positive rate than that in lymphoma, neuroblastoma, rhabdomyosarcoma, Ewing's sarcoma of bone and osteosarcoma. However, no expression of ICAM-1 was observed in fibrosarcoma, nephroblastoma and primitive neuroectodermal tumor patients. On the other hand, the expression rate of ICAM-1 was 55 in ALL, 65 in ANLL M1, M2, M3, and 50 in ANLL M4, M5. It is concluded that the expression of ICAM-1 in pediatric tumor and AL has variability. The ICAM-1 positive expression is observed in hepatoblastoma and ANLL M1, M2, M3 patients, whereas it is undetectable in fibrosarcoma, nephroblastoma and primitive neuroectodermal tumor patients.

Collision metastasis of breast and thyroid carcinoma to a single cervical lymph node: report of a case.

We herein report a rare case of collision lymph node metastases of breast and thyroid carcinomas. A 49-year-old female had undergone an extensively radical mastectomy of the right breast for inflammatory breast cancer at our hospital. Eleven months later, she presented with enlarged lymph nodes in her right lateral neck and multiple nodules in bilateral thyroid lobes. The patient underwent total thyroidectomy and radical dissection of the bilateral cervical lymph nodes. A histological examination showed multiple foci of papillary thyroid carcinoma (PTC) in the bilateral lobes. Surprisingly, concurrent metastases of breast carcinoma and PTC were shown in one of the lymph nodes from the right jugular region. This rare case of collision metastasis and the related literature are discussed.

IMP3 is a novel biomarker to predict metastasis and prognosis of tongue squamous cell carcinoma.

Insulinlike growth factor II mRNA-binding protein 3 (IMP3) is a newly identified mRNA-binding protein that is involved in embryogenesis and carcinogenesis of some malignant tumors. The aim of this study was to investigate the clinicopathologic significance of this protein in tongue squamous cell carcinoma (SCC). The expression of IMP3 in 65 samples of tongue SCC and 27 cases of oral leukoplakia (OL) was evaluated by immunohistochemistry. These expression levels were correlated with clinical and pathologic features as well as death from tongue SCC. Weak immunohistochemical stain for IMP3 was identified in all 19 cases of OL with mild dysplasia, and no immunohistochemical reactivity was found in 8 cases of OL without dysplasia. Positive immunohistochemical stain for IMP3 was identified in 50 cases (77%) of SCC; among them, weak staining was identified in 33 cases (51%) and intermediate staining in 17 cases (26%). To compare the expression of IMP3 in tongue SCC and OL, stronger immunohistochemical reactivity was found in tongue SCC (P < 0.05). Stronger expression of IMP3 was found to be associated with lymphoid metastasis (P < 0.05) and patient poor outcome (median survival time of 40 months in the negative and weak expression group vs 10 months in the intermediate expression group; P < 0.05). This study suggests that the increase in IMP3 expression in tongue leukopathia and SCCs may play a role in the carcinogenesis and tumor metastasis of tongue SCCs. Insulinlike growth factor II mRNA-binding protein 3 could be a novel prognostic indicator for patients with tongue SCCs.

Expression of Cyr61 in primary salivary adenoid cystic carcinoma and its relation to Ki-67 and prognosis.

Cysteine-rich protein 61 (Cyr61) selectively binds heparin and insulin-like growth factors and mediates a variety of biological actions, including cell adhesion, differentiation, proliferation, migration, angiogenesis, and tumorigenesis. Cyr61 is also a prognostic factor for tumor progression and survival of individuals with various types of tumors. This study investigated the relationship between the expression level of Cyr61 and clinicopathological features, as well as the prognostic significance of Cyr61 expression in human salivary adenoid cystic carcinoma (SACC). The expression of Cyr61 and Ki-67, a cell-proliferation marker, was examined immunohistochemically in paraffin embedded tissue specimens from 60 SACC patients who underwent radical surgery between 1995 and 2004. A chi-square test was used to investigate the relationship between Cyr61 and Ki-67 expression and clinicopathological features. Survival analysis was performed to determine the prognostic significance of Cyr61 expression. Cyr61 expression was observed in 39 cases (39/60, 65%) of SACC, and Cyr61 expression was positively correlated with Ki-67 expression (P=0.002). A high expression of Cyr61 was significantly associated with solid subtype, perineural invasion, vascular invasion or cancer embolus, advanced stage, recurrence, and metastasis (P<0.05). The survival rate of patients with high expression of Cyr61 or Ki67 was significantly lower than that of patients with low expression. Multivariate Cox's proportional hazards analysis showed that vascular invasion, TNM stage, recurrence, distant metastasis, Ki-67 expression, and Cyr61 expression were independent prognostic factors of overall survival (P<0.05). Cyr61 expression is significantly correlated with Ki-67 expression and may have potential value in screening high-risk cases for recurrence and metastasis, as well as identifying poor prognosis in SACC patients.

[Relationship of epidermal growth factor receptor gene mutations, clinicopathologic features and prognosis in patients with non-small cell lung cancer].

OBJECTIVE: To investigate epidermal growth factor receptor (EGFR) gene mutations in exons 19 and 21 of patients with non-small cell lung cancer (NSCLC) and to analyze the relationship of EGFR mutations with clinicopathological features and prognosis. METHODS: The EGFR gene exons 19 and 21 of paraffin-embedded tumor tissue were amplified by PCR, followed by direct sequencing in 282 surgically-removed specimens of NSCLC. The relationship of EGFR gene mutations in NSCLC with clinicopathological features and prognosis were analyzed. RESULTS: EGFR mutations were detected in 120 of 282 (42.6%) patients with NSCLC. There were 61 cases of the mutations in exon 19 and 66 cases of the mutations in exon 21, including 7 cases of the mutations both in exons 19 and 21. Mutations were more frequently observed in women (55.2%, 53/96) than in men (36.0%, 67/186), in 51 to 60-years-old (51.3%, 39/76) than ≤50-years-old (30.4%, 21/69) and >60-years-old (43.8%, 60/137), in non-smokers (54.3%, 69/127) than smokers (32.9%, 51/155), there was negative correlation of EGFR mutations with smoking status (P=0.000, rs=-0.216). EGFR mutations were more frequently observed in adenocarcinomas (47.8%, 64/134), bronchiolo-alveolar carcinomas (73.0%, 27/37), adenosquamous carcinomas (7/9) than squamous cell carcinomas (23.6%, 17/72) and other types (16.7%, 5/30). The EGFR mutation rate in the well differentiated, the middle differentiated, the poorly differentiated and the undifferentiated was 55.7% (68/122), 50.8% (30/59), 22.7% (17/75), 19.2% (5/26) respectively, the incidences of EGFR mutations decreased with the degrading of differentiation, there was positive correlation of EGFR mutations with differentiation of lung cancer (P=0.000, rs=0.296). The patients with EGFR mutations had better prognosis than those with wild-type EGFR (P=0.027). There was no association of EGFR mutations with clinical TNM stage. CONCLUSIONS: EGFR mutations occur frequently in females, non-smokers and adenocarcinomas, bronchioloalveolar carcinomas, and adenosquamous carcinomas. The patients with EGFR mutations have better prognosis. The results may offer a practical approach to select the patients who may benefit from anti-EGFR target therapy.

Expressions of connexin 32 and 26 and their correlation to prognosis of non-small cell lung cancer.

BACKGROUND AND OBJECTIVE: Gap junction intercellular communication (GJIC) plays an important role in regulating homeostasis and differentiation in many tissues. Connexins are gap junction proteins, whose expressions directly affect the function of GJIC. This study was to investigate expressions of connexin 32 and 26 proteins in non-small cell lung cancer (NSCLC), and their correlation to clinicopathological characters of NSCLC. METHODS: Immunohistochemistry was applied to detect expressions of connexin 32 and 26 in 77 NSCLC tissues. Correlations of connexin 32 and 26 expressions to smoking, tumor size, histological type, the degree of differentiation, lymph node metastasis and prognosis were analyzed. RESULTS: The positive rates of connexin 32 and 26 were 51.9% and 40.3% in the 77 samples, which were significantly higher than 20.3% and 30.5% in alveolar epithelium (p = 0.000, r = -0.322; p = 0.013, r = -0.215). Positive expression of connexion 32 was positively correlated with the differentiation degree of NSCLC tissues (p = 0.010, r = 0.345). The one- to five-year survival rates were higher in patients with positive connexion 32 expression than those without (p = 0.005). Moreover, the positive rate of connexin 26 was not correlated to smoking, tumor size, histological type, the degree of differentiation, lymph node metastasis and the postoperative survival time (p > 0.05). CONCLUSIONS: Expression of connexin 32 is closely correlated to the differentiation of NSCLC and affects the prognosis of NSCLC patients. Increasing the expression of connexin 32 may improve the prognosis of NSCLC.

[Expression and significance of CD44(+)ESA(+)CD24(-/low), stem cell markers for breast cancer, in non-small-cell lung carcinoma].

BACKGROUND & OBJECTIVE: Stem cells are found in all human tissues, while tumor stem cells (TSCs) are also detected in tumors. TSC of breast cancer has been separated and its markers have been affirmed. However, TSC of lung carcinoma has not been separated yet. This study was to investigate the expression and significance of stem cell markers for breast cancer (CD44(+)ESA(+)CD24(-/low)) in non-small-cell lung carcinoma (NSCLC). METHODS: Expressions of CD44,ESA and CD24 of tumor tissues in 77 cases of NSCLC patients were detected using immunohistochemistry. The correlations of the expression of the makers to tumor size, smoking, histological type, differentiation, lymphoid metastasis, and prognosis were analyzed. RESULTS: The expressive rates of CD44,ESA and CD24 were 63.6%, 66.2% and 7.8%, respectively in 77 NSCLC tissues. CD44-positive expression was significantly higher in poorly differentiated and undifferentiated group than in well differentiated group. ESA-positive expression was significantly higher in well differentiated group than in poorly differentiated and undifferentiated group. The ESA positivity was significantly higher in glandular carcinoma than in squamous carcinoma. The expressive rate of CD44(+)ESA(+)CD24(-/low) in 77 cases of NSCLC was 36.4%. No correlations were found in the expression of CD44(+)ESA(+)CD24(-/low) to smoking, tumor size, histological type, differentiation, lymphoid metastasis and prognosis (P>0.05). CONCLUSION: Expressions of stem cell markers for breast cancer (CD44(+)ESA(+)CD24(-/low)) are not associated with tumor size, histological type, differentiation, lymphoid metastasis and prognosis of NSCLC.

[Expression and clinical significance of vascular endothelial growth factor C and vascular endothelial growth factor receptor 3 in non-small cell lung carcinoma].

BACKGROUND & OBJECTIVE: Vascular endothelial growth factor C (VEGF-C) and vascular endothelial growth factor receptor 3 (VEGFR-3) play important roles in lymphangiogenesis of malignant tumors; their expression are closely related to lymphatic metastasis of malignant tumors. This study was designed to investigate the expression and clinical significance of VEGF-C and VEGFR-3 in non-small cell lung cancer (NSCLC). METHODS: The expression of VEGF-C and VEGFR-3 in 77 specimens of NSCLC were detected by immunohistochemistry; their correlations to lymphatic vessel density (LVD), tumor size, histological type, differentiation, lymphatic metastasis, clinical recurrence, and survival time of the patients were analyzed. RESULTS: Positive rate of VEGF-C was 58% in NSCLC, and that of VEGFR-3 was 42%. The expression of VEGF-C protein was negatively correlated with the differentiation of NSCLC (r=-0.32, P=0.018). The expression of VEGF-C and VEGFR-3 were related to lymphatic metastasis, LVD, tumor size, and survival time of the patients. The expression of VEGF-C was positively related with that of VEGFR-3 (r=0.23, P=0.045). CONCLUSION: The expression of VEGF-C and VEGFR-3 are closely related with lymphatic metastasis and prognosis of NSCLC; their high expression indicate high risk of lymphatic metastasis and poor prognosis.

Clinicopathological significance of expression of paxillin, syndecan-1 and EMMPRIN in hepatocellular carcinoma.

AIM: To evaluate the relationship of expression of paxillin, syndecan-1 and EMMPRIN proteins with clinicopathological features in hepatocellular carcinoma (HCC). METHODS: Fifty-one patients who underwent HCC resection were recruited in the study. Paxillin, syndecan-1 and EMMPRIN proteins in HCC tissues were detected with immunohistochemical staining. RESULTS: Of 51 cases of HCC, 23 (45%) exhibited paxillin protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 24 (57%) exhibited positive expression. Positive paxillin protein expression was associated with low differentiation (r = 0.406, P = 0.004), with the presence of portal vein thrombosis (r = 0.325, P = 0.021), with extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51 cases of HCC, 28 (55%) exhibited syndecan-1 protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 23 (55%) exhibited positive expression. Positive snydecan-1 protein expression was associated with well differentiation (r = 0.491, P = 0.001), with no extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51 cases of HCC, 28 (55%) exhibited EMMPRIN protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 21 (50%) exhibited positive expression. Expression of EMMPRIN protein was not associated with serum AFP level, HBsAg status, presence of microsatellite nodule, tumor size, presence of cirrhosis and necrosis, differentiation, presence of portal vein thrombosis, extra-hepatic metastasis, disease-free survival and overall survival (P>0.05). Expression of paxillin protein was correlated conversely with the expression of syndecan-1 protein in HCC (r = -0.366, P = 0.010). CONCLUSION: Expression of paxillin and syndecan-1 proteins in HCC may affect its invasive and metastatic ability of the tumor. There may be a converse correlation between the expression of paxillin and syndecan-1 protein in HCC. Expression of EMMPRIN protein may be detected in HCC, but it may play little role in the invasion and metastasis of HCC.

[Expression and significance of nitric oxide synthase mRNAs and proteins in giant cell tumors of bone].

BACKGROUND & OBJECTIVE: Recent studies have indicated that nitric oxide (NO) plays an important role in carcinogenesis and tumor progression. Activity of nitric oxide synthase (NOS) has been detected in normal bone cell lines. There was no report about relation of expression of NOS in giant cell tumors(GCT) of bone with its pathological grading and tumor recurrence. This study was designed to investigate the relationship amoung expression of NOS mRNAs, NOS proteins, pathological grading and tumor recurrence. METHODS: In situ hybridization (ISH) with cDNA probe was used to determine 14 frozen GCT specimens for constitutive NOS(cNOS) mRNA and inducible NOS(iNOS) mRNA. Immunohistochemical (IHC) staining with multiclonal antibodies was used to determine 42 paraffin-embedded GCT specimens for protein expression of NOS1, NOS2, and NOS3. RESULTS: (1)In 14 frozen GCT specimens, the positive expression rates of cNOS and iNOS mRNAs of multinuclear giant cells (MGC) were 78.6% and 57.1%; the positive expression rates of cNOS and iNOS mRNAs in mononuclear cells (MC) were both 35.7%. (2)The positive expression rate of cNOS mRNA in MGC of groups grading II and III was significantly higher than that of group grading I (P=0.008). (3) In 42 paraffin-embedded GCT specimens, the positive expression rates of NOS1, NOS2, and NOS3 protein were 85.7%, 59.5%, and 31.0% in MGC, 54.8%, 28.6%, and 14.3% in MC, respectively. (4)The positive expression rate of NOS1 protein in MC of groups grading II, III was significantly higher than that of group grading I (P=0.006). (5)The positive expression rate of NOS1 protein in MC of the recurrent group was significantly higher than that of the non-recurrent group (P=0.018). The positive expression rate of NOS3 protein in MGC of recurrent group was significantly higher than that of the non-recurrent group (P=0.041). CONCLUSION: The expression of NOS in GCT,especially cNOS in MC, is closely related to the pathological grading and the recurrence of GCT.