EMP1, a member of a new family of antiproliferative genes in breast carcinoma

This study aimed to analyze the expression, clinical significance of epithelial membrane protein 1 (EMP1) in breast carcinoma and the biological effect in its cell line by EMP1 overexpression. Immunohistochemistry and western blot were used to analyze EMP1 protein expression in 67 cases of breast cancer and 35 cases of normal tissues to study the relationship between EMP1 expression and clinical factors. Quantitative real-time RT-PCR and western blot were used to detect the mRNA level and protein of EMP1. MTT assay, migration and invasion assays were also conducted as to the influence of the upregulated expression of EMP1 that might be found on MCF-7 cell biological effect. The relative amount of EMP1 protein in breast cancer tissue was found to be significantly lower than in normal tissues (P?<?0.05). The level of EMP1 protein expression was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P?<?0.05). Meanwhile, loss of EMP1 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result shown that MCF-7 cell transfected EMP1 had a lower survival fraction, higher cell apoptosis, significant decrease in migration and invasion, higher caspase-9, and lower VEGFC protein expression compared with MCF-7 cell untransfected EMP1 (P?<?0.05). EMP1 expression decreased in breast cancer and correlated significantly with lymph node metastasis, clinic stage, histological grade, and poor overall survival, T stages, suggesting that EMP1 may play important roles as a negative regulator to breast cancer MCF-7 cell by regulating the expression of caspase 9 and VEGFC protein.     

Filamin A regulates MMP-9 expression and suppresses prostate cancer cell migration and invasion

This study aims to analyze the expression and clinical significance of Filamin A (FLNA) in prostate carcinoma and the biological effect in its cell line by FLNA overexpression. Immunohistochemistry and Western blot were used to analyze FLNA protein expression in 68 cases of prostate cancer and 37 cases of normal tissues to study the influence of the upregulated expression of FLNA that might be found on PC-3 cell biological effect. In the immunohistochemical analysis, the level of FLNA protein expression was found to be significantly lower in prostate cancer tissue than in normal tissues (P?<?0.05). In the Western blot analysis, the relative amount of FLNA protein in prostate cancer tissue was found to be significantly lower than in normal tissues (P?<?0.05). The level of FLNA protein expression was not correlated with age and PSA concentration (P?>?0.05), but it was correlated with T stages, lymph node metastasis, clinic stage, and Gleason score (P?<?0.05). The result of biological function showed that PC-3 cell transfected FLNA had a lower survival fraction, a significant decrease in migration and invasion, and a lower matrix metallopeptidase 9 (MMP-9) protein expression compared with PC-3 cell untransfected FLNA (P?<?0.05). FLNA expression decreased in prostate cancer and correlated significantly with T stages, lymph node metastasis, clinic stage, and Gleason score, suggesting that FLNA may play important roles as a negative regulator to prostate cancer PC-3 cell by promoting the degradation of MMP-9.     

CCNG2 expression is downregulated in colorectal carcinoma and its clinical significance

This study aimed to analyze the expression, clinical significance of cyclin G2 (CCNG2) in colorectal carcinoma, and the biological effect in its cell line by CCNG2 overexpression. Immunohistochemistry and Western blot were used to analyze CCNG2 protein expression in colorectal cancer and to study the influence of the upregulated expression of CCNG2 that might be found on SW480 cell biological effect. We found that the level of CCNG2 protein expression was significantly lower in colorectal cancer tissue than normal tissues (P?<?0.05). The level of CCNG2 was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P?<?0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result of biological function has shown that SW480 cell-transfected CCNG2 had a lower survival fraction, higher percentage of the G0/G1 phases, and lower CDK2 protein expression compared with SW480 cell-untransfected CCNG2 (P?<?0.05).     

Change in expression of cyclin G2 in kidney cancer cell and its significance

This study aims to analyze the expression and clinical significance of cyclin G2 (CCNG2) in kidney carcinoma, and the biological effect in its cell line by CCNG2 overexpression. Immunohistochemistry and western blot were used to analyze CCNG2 protein expression in 63 cases of kidney cancer and normal tissues to study the relationship between CCNG2 expression and clinical factors. CCNG2 lentiviral vector and empty vector were respectively transfected into kidney ACHN cell line. During immunohistochemistry, the level of CCNG2 protein expression was found to be significantly lower in kidney cancer tissue than normal tissues (P?<?0.05). After Western blot, the relative amount of CCNG2 protein in kidney cancer tissue was respectively found to be significantly lower than in normal tissues (P?<?0.05). The level of CCNG2 protein expression was not correlated with gender, age, tumor size, and pathological types (P?>?0.05), but it was correlated with lymph node metastasis, clinic stage, and histological grade (P?<?0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result of biological function show that ACHN cell-transfected CCNG2 had a lower survival fraction, higher percentage of the G0/G1 phases, and lower CDK2 protein expression compared with ACHN cell-untransfected CCNG2 (P?<?0.05). CCNG2 expression decreased in kidney cancer and correlated significantly with lymph node metastasis, clinical stage, histological grade, and poor overall survival, suggesting that CCNG2 may play important roles as a negative regulator to kidney cancer ACHN cell by promoting degradation of CDK2.     

Decreased expression of CCNG2 is significantly linked to the malignant transformation of gastric carcinoma

This study aimed to analyze the expression, clinical significance of cyclin G2 (CCNG2) in gastric carcinoma, and the biological effect in its cell line by CCNG2 overexpression. Immunohistochemistry and western blot were used to analyze CCNG2 protein expression in 87 cases of gastric cancer and normal tissues to study on the relationship between CCNG2 expression and clinical factors. CCNG2 lentiviral vector was transfected into gastric SGC-7901 cell line. RT-PCR and western blot were used to detect the mRNA level and protein of CCNG2. MTT assay and cell cycle were also conducted as to the influence of the upregulated expression of CCNG2 that might be found on SGC-7901 cell biological effect. Immunohistochemically, the level of CCNG2 protein expression was found to be significantly lower in gastric cancer tissue than in normal tissues (P?<?0.05). Western blot shows that the relative amount of CCNG2 protein in gastric cancer tissue was found to be significantly lower than in normal tissues (P?<?0.05). The level of CCNG2 protein expression was correlated with T stages, lymph node metastasis, clinical stage, and histological grade (P?<?0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result of biological function showed that SGC-7901 cell-transfected CCNG2 had a lower survival fraction, higher percentage of the G0/G1 phases, and lower CDK2 protein expression compared with SGC-7901 cell untransfected CCNG2 (P?<?0.05). CCNG2 expression decreased in gastric cancer and correlated significantly T stages, lymph node metastasis, clinical stage, histological grade, and poor overall survival, suggesting that CCNG2 may play important roles as a negative regulator to gastric cancer cell by promoting degradation of CDK2.     

BTG1 expression correlates with the pathogenesis and progression of breast carcinomas

This study aimed to analyze the expression, clinical significance of B cell translocation gene 1 (BTG1) in breast carcinoma and the biological effect in its cell line by BTG1 overexpression. Immunohistochemistry and western blot were used to analyze BTG1 protein expression in 72 cases of breast cancer and 36 cases of normal tissues to study the relationship between BTG1 expression and clinical factors. Recombinant lentiviral vector was constructed to over-express EMP-1 and then infect breast cancer MCF-7 cell line. Quantitative real-time RT-PCR (qRT-PCR) and western blot were used to detect the mRNA level and protein of BTG1. MTT assay, cell apoptosis, cell cycles, migration and invasion assays were also conducted as to the influence of the upregulated expression of BTG1 that might be found on MCF-7 cells biological effect. The level of BTG1 protein expression was found to be significantly lower in breast cancer tissue than normal tissues (P?<?0.05). Decreased expression of BTG1 was significantly correlated with tumor invasion, lymph node metastasis, clinic stage and histological grade of patients with breast cancer (P?<?0.05). Meanwhile, loss of BTG1 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result of biological function shown that MCF-7 cell transfected BTG1 had a lower survival fraction, higher percentage of the G0/G1 phases, higher cell apoptosis, significant decrease in migration and invasion, and lower CyclinD1, Bcl-2, and MMP-9 protein expression compared with MCF-7 cell untransfected BTG1 (P?<?0.05). BTG1 expression decreased in breast cancer and correlated significantly lymph node metastasis, clinic stage, histological grade, poor overall survival, proliferation, and metastasis in breast cancer cell by regulating CyclinD1, Bcl-2, and MMP-9 protein expression, suggesting that BTG1 may play important roles as a negative regulator to breast cancer cell.     

The expression of BTG1 is downregulated in NSCLC and possibly associated with tumor metastasis

This study aimed to analyze the expression, clinical significance of B cell translocation gene 1 (BTG1) in nonsmall cell lung cancer (NSCLC) and the biological effect in its cell line by BTG1 overexpression. Immunohistochemistry and western blot were used to analyze BTG1 protein expression in 82 cases of NSCLC and 38 cases of normal tissues to study the relationship between BTG1 expression and clinical factors. Recombinant lentiviral vector was constructed to overexpress EMP-1 and then infect NSCLC H1299 cell line. Quantitative real-time RT-PCR and western blot were used to detect the mRNA level and protein of BTG1. 3-[4,5-dimethylthiazol -2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, cell apoptosis, cell cycles, and migration and invasion assays were also conducted as to the influence of the upregulated expression of BTG1 that might be found on H1299 cells biological effect. The level of BTG1 protein expression was found to be significantly lower in NSCLC tissue than normal tissues (P?<?0.05). Decreased expression of BTG1 was significantly correlated with lymph node metastasis, clinic stage, and histological grade of patients with NSCLC (P?<?0.05). Meanwhile, loss of BTG1 expression correlated significantly with poor overall survival time by Kaplan–Meier analysis (P?<?0.05). The result of biological function show that H1299 cell transfected BTG1 had a lower survival fraction; higher percentage of the G0/G1 phases; higher cell apoptosis; significant decrease in migration and invasion; and lower CyclinD1, Bcl-2, and MMP-9 protein expression compared with H1299 cell untransfected BTG1 (P?<?0.05). BTG1 expression decreased in NSCLC and correlated significantly with lymph node metastasis; clinical stage; histological grade; poor overall survival; cell proliferation; cell cycles; cell apoptosis; and migration and invasion in NSCLC cell by regulating CyclinD1, Bcl-2, and MMP-9 protein expression, suggesting that BTG1 may play important roles as a negative regulator to NSCLC cell.     

鼻咽癌中EMP1蛋白的表达及其临床意义

目的： 探讨上皮膜蛋白1(EMP1)在鼻咽癌中的表达及其临床意义。方法：采用免疫组织化学方法和Western blot检测75例鼻咽癌组织及距癌组织边缘2 cm以上的镜下未见癌浸润的正常鼻咽组织中EMP1蛋白的表达情况,并分析其与临床病理指标的关系。结果：免疫组织化学结果表明,EMP1蛋白在鼻咽癌组织及正常鼻咽组织中的表达阳性率分别为33.3%和74.2%,前者显著低于后者(P＜0.05)。Western blot结果表明,EMP1蛋白在鼻咽癌组织的相对表达量较正常鼻咽组织明显降低(P＜0.05)。EMP1蛋白表达与鼻咽癌T分期、有无淋巴结转移、临床分期以及细胞分化程度有关(P＜0.05)。结论：EMP1低表达可能促进了鼻咽癌的发病过程,在鼻咽癌的发生发展过程中具有重要作用。     

Role of JNK signaling pathway in sensitivity to radiotherapy of nasopharyngeal carcinoma

Objective

The aim of the study was to investigate the effect of c-Jun N-terminal protein kinase (JNK) signaling pathway on influencing the sensitivity to radiotherapy of human nasopharyngeal carcinoma CNE cells.

Methods

Human nasopharyngeal carcinoma CNE multicellular spheroids (MCS) were constructed with three dimensional cell culture methods. Western blot was employed to analyze the activity of JNK signaling pathway in MCS after X-ray irradiation, and the expression of caspase-3 protein before and after using SP600125 (a special inhibitor of JNK). X-ray induced cell apoptosis in MCS before and after treated with SP600125 were detected by TUNEL.

Results

The level of JNK phosphorylation in MCS was a dynamic course after radiation, and there was a phosphorylation peaks at 2 h later, the apoptotic rate of MCS (P < 0.05) and the expression of caspase-3 protein (P < 0.05) were significantly increased after treated with SP600125.

Conclusion

The transient activation of JNK played a important role in sensitivity to radiotherapy of CNE MCS via mediating survival signals, blocking this pathway accelerate cell apoptosis, which may be related to the increased expression of caspase-3.     

NOB1 in Non-small-cell Lung Cancer: Expression Profile and Clinical Significance

Nin one binding (NOB1) gene has been reported up-regulated in several types of cancer. The aim of this study was to investigate the expression profile of NOB1 in non-small-cell lung cancer (NSCLC) and assess the clinical significance. qRT-PCR was used in the detection of NOB1 mRNA expression both in NSCLC tissue and in adjacent normal lung tissue. Western blot analysis and immunohistochemistry were used in the detection of NOB1 protein expression. The clinicopathological implications of NOB1 were analyzed statistically. It was confirmed by RT-qPCR that expression of NOB1 mRNA in NSCLC cells was higher than in human lung cells (P?<?0.05), and NOB1 mRNA was also over-expressed in NSCLC tissue when compared with adjacent tissue and normal lung tissue (P?<?0.05). Western blot analysis showed that NOB1 protein was significant increased in NSCLC cell lines compared with human lung cell line. Western blot analysis and immunohistochemistry showed that NOB1 protein was significant increased in NSCLC tissue compared with adjacent tissue and normal lung tissue (P?<?0.05). There were significant associations between NOB1 expression and TNM stage, lymph node metastasis, and histopathological grade (P?<?0.05), but not gender, age, smoke, or tumor diameter (P?>?0.05). Our results suggest that enhanced expression of NOB1 gene plays an important role in the occurrence and development of NSCLC. NOB1 may be a potential therapeutic target in NSCLC.     

ARID1A: a potential prognostic factor for breast cancer

The aim of this study is to explore the expression of BAF250a protein in breast cancer and its association with the clinical and pathological characteristics and prognosis of breast cancer. The expression status of BAF250a protein was detected by Western blot analysis and immunohistochemical staining. The relationship between BAF250a proteins and clinicopathological parameters in 496 breast cancer specimens was analyzed. Western blot analysis showed that BAF250a protein had a lower expression in breast cancer specimens than in matched normal breast tissue (104.38?±?11.65 vs. 55.94?±?10.27; P?=?0.004, t test). Among the 496 enrolled breast cancer patients, BAF250a protein expression was absent in 324 (65.3 %). Universal and multiple analyses indicated that BAF250a protein expression loss was significantly related to histological grade, metastatic nodes, tumor node metastasis (TNM) stage, and the expression of estrogen receptor (ER), progesterone receptor (PR), c-erbB-2, and p53 (all P?<?0.05). For TNM stage, rank correlation coefficients were 0.199 and 0.191, respectively, (P?<?0.05), but BAF250a protein deletion had either a positive or a negative correlation with ER, PR, c-erbB-2, and p53 protein expression (correlation coefficients were 0.231, 0.207, ?0.098, ?0.128; P?<?0.05). Analysis of survival rates showed that the patients with BAF250a protein expression attained a significantly better postoperative disease-specific survival than those with BAF250a protein deletion (88.4 vs. 79.0 %; P?=?0.003). In the Cox regression test, BAF250a protein deletion was detected as an independent prognostic factor (P?=?0.014). BAF250a protein might be a new potential target for breast cancer treatment.     

Expression of p114RhoGEF predicts lymph node metastasis and poor survival of squamous-cell lung carcinoma patients

The Rho-specific guanine nucleotide exchanging factor p114RhoGEF is involved in RhoA activation and cell motility. Previous studies suggest that altered expression of p114RhoGEF could contribute to cancer progression. We investigated an association of p114RhoGEF expression with progression and prognosis of non-small cell lung cancer (NSCLC). Immunohistochemistry was performed to detect p114RhoGEF expression in 105 NSCLC (34 adenocarcinoma and 71 squamous-cell carcinoma) and 32 normal lung tissues. We found that p114RhoGEF expression was upregulated in squamous-cell lung carcinoma and that p114RhoGEF expression was significantly higher in squamous-cell carcinoma than in adenocarcinoma or normal tissues (P?<?0.05, both). Expression of p114RhoGEF protein was significantly associated with lymph node metastasis of lung cancer (P?<?0.05), but not with patients’ age, gender, tumor size, differentiation, or stage. Expression of p114RhoGEF protein was also associated with poor overall and event-free survival of squamous-cell lung carcinoma patients (P?<?0.05). Taken together, p114RhoGEF expression may be useful in predicting progression and survival of squamous-cell lung carcinoma patients. A future study will investigate whether p114RhoGEF can serve as a novel therapeutic target in squamous-cell lung cancer.     

Clinical Significance of Mannose-Binding Lectin Expression in Thyroid Carcinoma Tissues

Mannose-binding lectin (MBL) plays an important role in the host defence against pathogens and carcinogenesis. This study aimed to analyze differential expression of MBL protein in thyroid cancer tissues and then to investigate the effects of rhMBL in thyroid cancer cells. Tissue specimens from 45 thyroid carcinoma patients and 45 adenoma patients were recruited for immunohistochemical analysis of MBL expression. Cell viability, apoptosis, RT-PCR and Western blot assays were used to detect changes in tumor cell viability, apoptosis, and gene expression, respectively, after treatment of thyroid cancer cells with rhMBL. MBL was differentially expressed in papillary thyroid carcinoma, adenoma, and the distant normal tissues (0.322?±?0.008, 0.227?±?0.003, and 0.113?±?0.003, respectively, P?<?0.05). MBL expression was associated with the advanced disease stage, histological grade, or lymph node metastasis in cancer patients (P?<?0.05). Moreover, rhMBL treatment of thyroid cancer cells reduced tumor cell viability but induced apoptosis in a dose- and time-dependent manner. rhMBL treatment also downregulated Bcl2 protein expression in thyroid cancer cells (P?<?0.05). In addition, expression p53 protein was increased in thyroid cancer cells after rhMBL treatment (P?<?0.05). The data from the current study demonstrate that MBL overexpression is associated with advanced thyroid carcinomas, and rhMBL treatment significantly reduced viability but induced apoptosis of thyroid cancer cell lines. Further studies will clarify whether overexpressed MBL in thyroid cancer tissues is functional.