Relationship between tumor metastasis suppressor gene KAI1 and the invasive and metastatic characteristics of osteosarcoma

AIM:To study the expression of metastasis suppressor gene KAI1 mRNA in osteosarcoma tissue and osteosarcoma cell lines,and the relationship between it and the biological behavior of the tumor cells.METHODS:RT-PCR was used to detect KAI1 mRNA in 18 cases of resected fresh osteosarcoma samples and three cultured osteosarcoma cell lines.The proliferative rate,the adhesive and invasive abilities of the 3 cell lines were detected.The results were treated by analysis system of images and analyzed with t test.RESULTS:The relative amount of KAI1 mRNA in osteosarcomas with lung metastasis was 0.80±0.50,while that was 1.48±0.64 in osteosarcomas without lung metastasis,the former was significantly lower than the latter (P<0.05).However,KAI1 mRNA had no corelation with the recurrence of osteosarcoma.The expression of KAI1 mRNA in U2-OS was highest (P<0.05),while the proliferation rate,the adhesive and invasive ability of U2-OS were the lowest among the 3 cell lines (P<0.01).CONCLUSION:The metastasis suppressor gene KAI1 might take part in influencing the lung metastasis of osteosarcoma,which might be caused by inhibiting the tumor cell proliferation,adhesion and invasion.     

Mortalin expression in cervical squamous-cell carcinoma and its effect on tumor metastasis

AIM: To investigate the significance of mortalin expression in clinical pathology of cervical squamous-cell carcinoma. METHODS: Immunofluorescence staining was used to detect the location of mortalin in human cervical squamous-cell carcinoma SiHa cells. The protein expression of mortalin was detected in 59 cases of normal cervical epithelial tissues and 93 cases of cervical squamous-cell carcinoma tissues by immunohistochemical staining, and its correlation with clinicopathological features of cervical squamous-cell carcinoma was also analyzed. MTT assay was used to evaluate the optimal concentration and dosing time of mortalin inhibitor MKT-077. After the protein expression of mortalin in SiHa cells was inhibited, wound-healing and migration assays were performed. The protein expression of epithelial-mesenchymal transition (EMT)-related molecules was determined by Western blot. RESULTS: Immunofluorescence staining showed that mortalin was located in the cytoplasm of SiHa cells. The positive rate and strongly positive rate of mortalin in the cervical squamous-cell carcinoma patients were 88.7% (55/62) and 61.3% (38/62), respectively, and they were significantly higher than those in normal cervical epithelial tissues (23.7% and 5.1%, P < 0.01). Additionally, mortalin expression was statistically correlated with the histological grade, clinical stage and lymph node metastasis. After inhibiting the expression of mortalin in the SiHa cells by MKT-077, the results of wound-healing and migration assays showed that the migration ability of SiHa cells was down-regulated. The protein expression of E-cadherin was up-regulated, and vimentin and Snail were significantly down-regulated. CONCLUSION: Mortalin over-expression is an effective biomarker for prediction of malignant potential and poor prognosis of cervical squamous-cell carcinoma.     

《中国蔬菜品种志》

本书由中国农业科学院蔬菜花卉研究所主编,已于2002年9月出版发行。全书分上、下卷,1～6章为上卷,包括根菜类、白菜类、芥菜类、甘蓝类、绿叶菜类及葱蒜类,计2263个品种,1347页;7～12章为下卷,包括瓜类、茄果类、豆类、薯芋类、水生蔬菜类和多年生蔬菜类,计2550个品种,1177页。入志的品种中,地方品种占     

Expression of HDAC6 protein in cervical carcinoma tissues and its clinical significance

AIM:To investigate the protein expression of histone deacetylase 6(HDAC6) in cervical carcinoma tissues and its clinical value. METHODS:The method of immunohistochemistry was used to detect the protein expression of HDAC6 in 63 cases of cervical carcinoma tissues, 38 cases of cervical intraepithelial neoplasia(CIN) tissues and 63 cases of normal cervical epithelial tissues. The relationships between the protein expression of HDAC6 and clinical pathological features were analyzed. The protein expression of HDAC6 in randomly selected 4 cases of cervical carcinoma tissues and paired normal cervical epithelial tissues was detected by Western blotting. RESULTS:Positive rates of HDAC6 protein expression in cervical carcinoma tissues were significantly higher than that in CIN tissues or normal cervical epithelial tissues, and there were obvious differences among the 3 groups(P<0.05). The protein expression of HDAC6 was not related to age and histological differentiation(P>0.05), but closely associated with clinical stages, invasive depth and lymph node metastasis(P<0.01 or P<0.05). Furthermore, the result of Western blotting demonstrated that the protein level of HDAC6 in cervical carcinoma tissues was markedly higher than that in normal cervical epithelial tissues. CONCLUSION:HDAC6 may be an important molecular marker for evaluating malignant degree and prognosis of cervical carcinoma.     

Abnormal expression and clinical significance of RASA1 in pancreatic carcinoma

AIM: To investigate the role of Ras p21 protein activator 1 (RASA1) in the development and progress of pancreatic cancer. METHODS: Different expression levels of RASA1 were measured by qRT-PCR and Western blotting in the pancreatic cancer cells Capan-2, CFPAC-1 and BxPC-3, and the pancreatic ductal cell line H6C7. Besides, the different expression levels between the pancreatic cancer and the pancreatic benign lesions, such as chronic pancreatitis or pancreatic cyst, were detected by the method of immunohistochemistry. The relationship between the clinicopathological feature and the RASA1 expression was analyzed. RESULTS: Both mRNA and protein expression levels of RASA1 decreased in pancreatic cancer cells compared with the ones in the pancreatic ductal cells (P<0.05). The protein level of RASA1 in the pancreatic cancer tissues was lower than that in the pancreatic benign lesion tissues (P<0.05). The pancreatic cancer samples with adjacent organ invasion had a significantly lower expression level of RASA1 than that in the pancreatic cancer samples limited in the pancreas (P<0.05). The expression levels of RASA1 were much higher in the cancers on stage I than the ones on stage II or Ⅲ (P<0.05). However, no relationship between the RASA1 expression level and the maximum diameter of cancer, the lymph node invasion and the survival time was observed. CONCLUSION: RASA1 plays an important role in the pancreatic cancer development as a potential tumor suppressor.     

Methylation and mRNA expression of TIG1 gene in esophageal squamous-cell carcinoma tissues

AIM: To investigate the expression and promoter methylation of tazarotene-induced gene-1 (TIG1) in esophageal squamous-cell carcinoma (ESCC) tissues. METHODS: The methods of methylation-specific PCR and real-time fluorescence quantitative PCR were applied to examine the methylation and mRNA expression of TIG1, respectively, in 43 cases of ESCC tissues, 20 cases of paracancerous tissues and 15 cases of normal tissues. RESULTS: The frequency of promoter methylation of TIG1 gene in ESCC tissues was 25.6% (11/43), which was significantly higher than that in the paracancerous tissues (5.0%, 1/20) and normal tissues (0/20). The hypermethylation of TIG1 gene in these tissues had no correlation with sex, age and clinical stage of the patients. However, it was correlated with the pathological stage (P<0.01) and lymph node metastasis (P<0.05). The mRNA expression of TIG1 in ESCC tissues was significantly lower than that in paracancerous tissues (P<0.05) and normal tissues (P<0.01). However, the expression level of TIG1 mRNA in methylated tissues was significantly lower than that in unmethylated tissues (P<0.01). CONCLUSION: Promoter methylation may be an important mechanism of TIG1 gene inactivation in ESCC, which was related to lymph node metastasis and TNM stage of esophageal carcinoma.     

Clinical significance of STMN1 expression in cervical cancer and effect of inhibition of its expression on viability and apoptosis of cervical cancer SiHa cells

AIM: To investigate the clinical significance of stathmin 1 (STMN1) expression in cervical cancer and the influence of its expression on the viability and apoptosis of cervical cancer cells. METHODS: Western blot was used to detect the protein expression of STMN1 in cervical cancer tissues, and the relationship between the expression and clinical characteristics of cervical cancer was analyzed. STMN1-siRNA was transfected into cervical squamous-cell carcinoma SiHa cells. The protein levels of STMN1, STAT3, p-STAT3 and survivin were determined by Western blot after transfection for 48 h. The cell viability was measured by MTT assay. The apoptosis was analyzed by flow cytometry. DCFH-DA probe was used to detect the level of reactive oxygen species (ROS). RESULTS: The protein expression of STMN1 in cervical cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The STMN1 protein expression level was not correlated with age and histological types of cervical cancer patients, but was related to clinical stage, histological differentiation and lymph node metastasis (P<0.01). Transfection with STMN1-siRNA significantly reduced the expression of STMN1 in SiHa cells. Compared with control group, the cell viability in STMN1-siRNA group was significantly decreased, the apoptotic rate and ROS content were increased, and the protein levels of p-STAT3 and survivin were down-regulated (P<0.01). However, no significant difference of the STAT3 protein level was observed between STMN1-siRNA group and control group. CONCLUSION: STMN1 is highly expressed in cervical cancer, and its expression is related to clinical stage, histological differentiation and lymph node metastasis. Inhibition of STMN1 expression reduces the viability and promotes apoptosis of cancer cells by down-regulating STAT3 signaling pathway.     

Expression and clinical significance of Bmi-1 in gastric carcinoma

AIM: To investigate the relationship between expression of Bmi-1 (B cell-specific MLV integration site-1) in gastric cancer and its clinicopathologic significance.METHODS: 146 surgical patients with gastric carcinoma were followed up at least 2 years.Expression of Bmi-1 protein was examined by immunohistochemistry in their archival paraffin embedded tissue specimens.RESULTS: The intensive positive rate of Bmi-1 expression in gastric cancer was 67.8% (99/146).Expression of Bmi-1 was highly correlated with tumor size,clinical stage,lymph node metastasis and T classification (P<0.05),but not with sex,age,tumor differentiation,etc.(P>0.05).The survival rate in the patients with Bmi-1 expression was much lower than that in those patients without Bmi-1 expression (P<0.01).Multivariate analysis indicated that Bmi-1 expression,T classification,lymph node metastasis,distant metastasis,tumor size and postoperative chemotherapy were all significantly prognostic factors of gastric carcinoma.CONCLUSION: Overexpression of Bmi-1 in patients with gastric carcinoma enhances the possibility of invasion and metastasis,implying a poor prognosis.Bmi-1 may serve as fairly a good prognostic factor to indicate biologic behavior and prognosis in gastric carcinoma.     

Expression of adhesion molecules in hepatocellurlar carcinoma and its clinical significance

AIM: To investigate the expression of adhesion molecules in hepatocellular carcinoma (HCC), and analyze its clinical significance. METHODS: The expressions of adhesion molecules of tumor tissues of 64 cases and adjacent tissues of 12 cases of HCC were detected with RT-PCR. RESULTS: ①The expression rates of E-cadherin, ICAM-1, CD44, CD44V, α₅, β₁ were 90.62%, 93.75%, 50.00%, 96.88%, 100%, 100%, respectively, and there was a significant difference between CD44 and other adhesion molecules. ②The expression level of E-cadherin, ICAM-1, CD44, CD_44V, α₅,β₁ in liver cancer tissues were 1.24±0.54, 0.96±0.37, 0.62±0.73, 0.86±0.33, 0.97±0.49, 1.41±0.24, respectively, and there was a significant difference between CD44 and E-cadherin, β₁. ③The expression level of E-cadherin and CD44 mRNA declined as HCC stage become higher, and there was a statistical difference in the expression level of CD44 mRNA between Ⅰ-Ⅱ stage and Ⅳ stage. The expression level of ICAM-1, α₅, β₁ had a trend to rise as HCC stage become higher, and there was a statistical difference in the expression level of ICAM-1 between Ⅰ-Ⅱ stage and Ⅳ stage. ④The expression level of ICAM-1,CD_44V, α₅, β₁ had positive correlation with tumor volume, tumor nodules, tumor metastasis, and had negative correlation with tumor encapsulation. E-cadherin and CD44 had negative correlation with tumor volume, tumor nodules, tumor metastasis, and had positive correlation with tumor encapsulation. All showed no significant correlation with the level of AFP , the degree of cirrhosis and the function of liver. CONCLUSION: There was a significant difference in the expression level of adhesion molecule mRNA in HCC, and their expression had Spearman correlation with each other. The expression level of adhesion molecule mRNA is associated with tumor volume, tumor nodules and tumor metastasis.     

Expression of glypican-3 in hepatocellular carcinoma and its clinical significance

AIM: To study the expression of glypican-3 in hepatocellular carcinoma (HCC) tissues and to clarify its clinical significance. METHODS: The expression of GPC3 was detected in 59 cases of HCC and their para-cancerous tissues, 10 cases of intrahepatic cholangiocellular carcinoma (ICC), 11 cases of cirrhotic tissues and 14 cases of normal liver tissues (around haemangioma) by RT-PCR and immunohistochemical staining. The survival curves were constructed using Kaplan-Meier method and evaluated using the log-rank test. In addition, the Cox proportional hazards regression model was established to identify the factors that were independently associated with disease-free survival (DFS). RESULTS: The mRNA expression of GPC3 in the HCC tissues was significantly higher than that in the para-cancerous tissues (83.1% vs 35.6%, χ²=27.53, P<0.01). The protein expression of GPC3 in the HCC tissue was also higher than that in the para-cancerous tissues (78.0% vs 33.2%, χ²=24.97, P<0.01). The expression of GPC3 in ICC tissues, liver cirrhosis tissues and normal liver tissues was undetectable. Kaplan-Meier survival analysis indicated that the GPC3(+)HCC patients had worse 1-year DFS than that of GPC3(-) patients (33.6% vs 72.7%, P<0.05). The HCC patients with para-cancerous GPC3(+) also had worse 1-year DFS than that of the para-cancerous GPC3(-) patients (23.5% vs 40.1%, P<0.05). The DFS rate decreased significantly as the expression intensity of GPC3 increased. The Cox regression model analysis indicated that AFP(+) (odd ratio=0.372, 95% confidence interval: 0.140-0.900, P<0.05), tumor size (odd ratio=5.215, 95% confidence interval: 1.737-15.656, P<0.01), para-cancerous tissue GPC3(+) (odd ratio=0.226, 95% confidence interval: 0.085-0.599, P<0.01) and the intensity of GPC3 expression in HCC tissue (odd ratio=1.946, 95% confidence interval: 1.080-3.507, P<0.05) were the independent risk factors linked to DFS of patients. CONCLUSION: GPC3 protein is highly expressed in the HCC tissues,but not in ICC, cirrhotic liver and normal liver tissues. The expression of GPC3 in para-cancerous tissues and the intensity of GPC3 expression in HCC tissues are the important independent risk factors linked to DFS of patients.     

The expression and significance of Apaf-1 in papillary thyroid carcinoma

AIM: To investigate the mRNA and protein expression of apoptotic protease-activating factor 1 (Apaf-1) in papillary thyroid carcinoma (PTC) and its relationship with cell proliferation.METHODS: The methods of real-time PCR, Western blot and immunohistochemistry were used to detect the mRNA and protein expression of Apaf-1 in PTC and tumor-adjacent tissues. The relationship between Apaf-1 expression and clinicopathological characteristics was analyzed. The effect of Apaf-1 on cell proliferation was verified by down-regulating the expression of Apaf-1 in CGTHW-3 cells.RESULTS: The mRNA and protein expression levels of Apaf-1 in the PTC tissue were significantly lower than those in the tumor-adjacent tissue (P<0.05). Down-regulation of Apaf-1 expression enhanced the proliferation of CGTHW-3 cells (P<0.05).CONCLUSION: The expression of Apaf-1 is low in PTC, and inhibition of its expression enhances the proliferation of CGTHW-3 cells. Apaf-1 may play a tumor suppressor role in PTC.     

Expression of SCD-1 in cervical carcinoma and effect of its down-regulation on proliferation and apoptosis of cervical carcinoma cells

AIM:To examine the expression of stearoyl-CoA desaturase-1 (SCD-1) in normal cervical tissues, cervical squamous cell carcinoma tissues, and cervical carcinoma cell lines HeLa, SiHa and CaSki, and to investigate the effect of down-regulation of SCD-1 on the proliferation and apoptosis of cervical carcinoma cells. METHODS:The expression of SCD-1 was detected by Western blotting in normal cervical tissues, cervical squamous cell carcinoma tissues, and cervical carcinoma cell lines HeLa, SiHa and CaSki. SCD-1 siRNA and control siRNA were utilized to transfect CaSki cells by Lipofectamine 2000, and SCD-1 protein level was determined by Western blotting after transfection. Furthermore, CCK-8 and flow cytometry were utilized to investigate the changes of cell proliferation and apoptosis after transfection with SCD-1 siRNA in CaSki cells. Subsequently, the activities of caspase-3 and caspase-9 were analyzed by Caspase-Glo3/7 and 9 detection kit after transfection with SCD-1 siRNA in CaSki cells. Finally, the protein expression of Bcl-2 and Bax was detected by Western blotting. RESULTS:The protein expression of SCD-1 in cervical squamous cell carcinoma tissues was significantly higher than that in normal cervical tissues, and the protein expression of SCD-1 in the 3 cervical carcinoma cell lines was obviously higher than that in normal cervical tissues, in which CaSki cells displayed the highest SCD-1 protein level. In addition, the protein expression of SCD-1 in SCD-1 siRNA group was significantly lower than that in untreated group and control siRNA group. Compared with untreated group and control siRNA group, the proliferation of CaSki cells was markedly inhibited in SCD-1 siRNA group. Early apoptotic rate in SCD-1 siRNA group was evidently higher than that in untreated group and control siRNA group. The activities of caspase-3 and caspase-9, and the level of Bax protein were significantly elevated, and the protein level of Bcl-2 was obviously reduced after transfection with SCD-1 siRNA in CaSki cells. CONCLUSION: SCD-1 may play an important role in the occurrence and development of cervical carcinoma, and its down-regulation, which mediates cell proliferation inhibition and apoptosis, may be tightly associated with the activities of caspase-3 and caspase-9, and the protein expression of Bcl-2 and Bax.     

Expression and clinical significance of SCNM1 in hepatitis B-related hepatocellular carcinoma

AIM: To investigate the expression of sodium channel modifier 1 (SCNM1) in hepatitis B-related hepatocellular carcinoma (HCC) and its relationship with clinicopathological features and prognosis.METHODS: The specimens were collected from 108 patients with hepatitis B-related HCC who were treated in the Third Affiliated Hospital of Sun Yat-sen University from January 2013 to December 2015. All patients signed the informed consent and met the requirements of medical ethics. The mRNA expression level of SCNM1 in hepatitis B-related HCC tissues and tumor-adjacent tissues was detected by RT-qPCR, and the relationship between the mRNA expression of SCNM1 and the clinicopathological characteristics of hepatocellular carcinoma was analyzed. The relationship between SCNM1 expression and the prognosis of the patients was analyzed by Kaplan-Meier plotter.RESULTS: The data from TCGA database, Human Protein Atlas database and Oncomine database showed that the expression of SCNM1 in hepatocellular carcinoma tissues was significantly higher than that in the normal liver tissues (P<0.01). SCNM1 was mainly distributed in the nucleus. The results of RT-qPCR showed that the median mRNA expression of SCNM1 in hepatitis B-related HCC tissues was significantly higher than that in the matched tumor-adjacent tissues (t=8.082, P<0.01). The mRNA expression of SCNM1 was correlated with cirrhosis, alanine aminotransferase and tumor size (P<0.05), but not with sex, age and tumor envelope. The total survi-val time of the HCC patients with high expression of SCNM1 was shorter than that of the patients with low expression of SCNM1 (HR=1.53, P=0.016), and that of the patients with hepatitis B-related HCC was even shorter (HR=2.41, P=0.015).CONCLUSION: SCNM1 is highly expressed in hepatitis B-related HCC and may play an important role in the development of hepatitis B-related HCC.     

Expression of nitric oxide synthase and its clinical significance in hepatic cellular carcinoma

AIM: To investigate the expression of nitric oxide synthase (NOS) and its clinical significance in hepatic cellular carcinoma (HCC). METHODS: The NOS1, NOS2 and NOS3 of 51 cases of HCC and 46 cases of liver tissue beside carcinoma (LTBC) were detected by immunohistochemistry. RESULTS: The expressive rates of NOS1, NOS2 and NOS3 in LTBC were significantly higher than those in HCC (P<0.01). The expressive rates of NOS1 in the recurrent group was significantly higher than that in the non-recurrent group (P<0.01). The expressive rate of NOS2 in the group without carcinoma embolus was significantly higher than that in the group with carcinoma embolus (P<0.05). The expressive rate of NOS3 in the recurrent group was significantly higher than that in the non-recurrent group (P<0.01). CONCLUSION: The expression of NOS1, NOS2 and NOS3 are closely related with the biological behaviors of HCC.     

Expression of ALEX1 in breast cancer and its clinical significance

AIM: To detect the expression of ALEX1 in the breast cancer tissues in order to verify whether ALEX1 has correlation with clinical pathological features in breast cancer.METHODS: Real-time PCR and immmunohistochemistry were applied to detect the expression of ALEX1 at mRNA and protein levels in the breast tissues. The statistical analysis were performed for determining the correlation with the level of ALEX1 and the clinical pathological features in breast cancer.RESULTS: The protein levels of ALEX1 in the breast cancer tissues were lower than that in the non-breast cancer tissues (P<0.01). The expression of ALEX1 had correlations with pathological grade, clinical stage, molecular type (P<0.05) but had no correlation with the patients' age, tumor size and tumor types in breast cancer. Furthermore, the result of real-time PCR showed that mRNA expression of ALEX1 was also significantly reduced in the breast cancer tissues (P<0.01).CONCLUSION: The expression of ALEX1 in the breast cancer tissues is lower than that in non-breast cancer tissues. The pathological grade and clinical stage in breast cancer are negatively correlated with the expression of ALEX1.     

Expression of KDM5B gene in breast cancer and its clinical significance

AIM:To investigate the expression of KDM5B gene in breast cancer tissues and its relationship with clinical data and prognosis of the patients. METHODS:Data sets of breast cancer were collected from The Cancer Genome Atlas (TCGA) database, and KDM5B mRNA expression profiles were downloaded. The mRNA expression of KDM5B in breast cancer tissues and adjacent tissues was detected by real-time PCR. The cases were divided into high expression group and low expression group according to the median expression of KDM5B, and the relationship with clinical data and case characteristics were analyzed. The relationship between KDM5B and prognosis of breast cancer patients was analyzed by Kaplan-Meier plotter. RESULTS:The expression of KDM5B in breast cancer tissues was significantly higher than that in normal breast tissues (P<0.01). In TCGA breast cancer data, the expression of KDM5B was significantly correlated with human epidermal growth factor receptor 2 (HER2), estrogen receptor (ER), age, histopathological type and lymph node metastasis (P<0.01), but not with progesterone receptor (PR), menopause and distant metastasis. The expression of KDM5B was significantly correlated with HER2, age and lymph node metastasis, but not with ER, PR, menopause, pathological type and distant metastasis. The higher the expression of KDM5B, the shorter the total survival time and the disease-free survival time of breast cancer patients. CONCLUSION:KDM5B is over-expressed in breast cancer tissues and correlated with prognosis of the patients. KDM5B expression is significantly correlated with HER2, age and lymph node metastasis. KDM5B may play an important role in the development of breast cancer.     

Expression of midkine and its clinical significance in transitional cell carcinoma of human urinary bladder

AIM: To investigate the expression of midkine in bladder transitional cell carcinoma and to analyze its relationship with the features of clinical pathology and prognosis.METHODS: The expressions of midkine protein in 50 cases of bladder transitional cell carcinoma samples were detected by SP immunohistochemical method using polyclonal antibodies against human midkine.Survival time of 40 cases was recorded.RESULTS: The protein expression of midkine was found in cytoplasm of tumor cells.The overall positive rate of midkine in 50 cases of bladder carcinoma was 90% (45/50).The positive degree of midkine showed a trend of increasing in grade and stage.There was statistically significant difference among them (P<0.05),but not with sex,age,treatment or tumor number and size (P>0.05).Patients with high expression of MK predicted a poor clinical outcome.CONCLUSION: Midkine is overexpressed in bladder transitional cell carcinoma than that in normal bladder.MK expression in bladder cancer is higher in less differentiated and deeper invaded cases,but it has no correlation with age,sex,treatment,tumor number and size.Patients with higher MK expression have shorter survival time than those with lower MK expression.