Prognostic Role of KRAS mRNA Expression in Breast Cancer / 한국유방암학회지

Effects of subchronic aluminum exposure on long-term potentiation and activities of RAS and extracellular regulated protein kinases in rats / 中华劳动卫生职业病杂志

Objective@#To study the effects of subchronic aluminum exposure on LTP and activities of RAS and ERK in rats in vivo.@*Methods@#24 Wistar rats were randomly divided into control group、low-dose group、medium-dose group and high-dose group, and received saline (control group) or Al (mal) 3 (15 μmol、kg、30 μmol、kg or 45 μmol/kg) via intraperitoneal injection (i.p.) for 8 weeks, respectively. The fEPSP in CA1 region were recorded by field potentiation technique in vivo and the hippocampal activities of RAS and ERK were examined by ELISA.@*Results@#The fEPSP amplitudes of the control group were 1.90±0.19, 1.64±0.15 and 1.54±0.08 at 1, 30 and 60 min after HFS, respectively. The fEPSP amplitudes of the low-dose group were 1.40±0.06 at 60 min, which represented a statistically significant decrease compared to the control group (P<0.05) ; these values at 30min and 60min dropped to 1.33±0.20 and 1.12±0.07 in the medium-dose group (P<0.05) and further decreased to 1.05±0.05 and 0.91±0.10 in the high-dose group (P<0.05) . And the activity dose-dependent decreases were observed both in RAS and ERK: compared with the control group and the low-dose group, the activities of RAS and ERK of the medium-dose and high-dose group significantly decreased (P<0.05) and compared with the medium-dose group, the activities of the high-dose group statistically dropped (P<0.05) .@*Conclusion@#RAS and ERK may be related to the suppression of LTP by subchronic aluminum exposure and the RAS-MAPK transduction pathway may be involved in the damage of learning and memory induced by aluminum.

Significance and expression of PAX8,PAX2,p53 and RAS in ovary and fallopian tubes to origin of ovarian high grade serous carcinoma / 中华妇产科杂志

Objective To explore the origin of ovarian high grade serous carcinoma(HGSC) through analysing the expression and significance of PAX8,PAX2,p53 and RAS in the ovary and fallopian tube of different types and grades of serous carcinoma. Methods A total of 44 cases tissue samples of ovarian tumor including 34 malignant ovarian tumor and 10 normal normal tissue (as control group) were collected from the admitted patients in Affiliated Tumor Hospital of Guangxi Medical University from January 2015 to January 2016. Fallopian tube tissues were segmented in accordance with the fimbria, ampulla, isthmus and the corresponding ovarian tissues were by the side. There were 34 cases of patients with ovarian cancer including 29 cases of epithelial ovarian cancer (27 serous carcinoma, 1 mucinous carcinoma,1 endometrioid adenocarcinoma)and 5 non-epithelial ovarian cancer(sex cord-interstitial tumor). Among 27 cases of patients with ovarian serous cancer,there were 23 HGSC and 4 low-grade ovarian serous cancer (LGSC). One hundred fifty-three cases of samples were diagnosed as ovarian serous cancer by Shandong University Affiliated Qilu Hospital from 2005 to 2013 and these samples were made tissue microarray.(1)To analyze the expression and differences of PAX8,PAX2,p53 and RAS in the above tissues and tissue microarray from ovarian and tubal of HGSC and control women by immunohistochemistry methods.(2)To compare the expression levels of PAX8,PAX2,p53 and RAS in ovarian and fallopian tubes of ovarian cancer patients with different pathological types. (3) To analyze the correlations of tubal and ovarian tissue in PAX8,PAX2,p53 and RAS expression of HGSC.(4)To analyze the factors of the prognosis of ovarian serous cancer in tissue microarray by single factor analysis method. Results (1)PAX8,PAX2, p53 and RAS expression was negative in normal ovarian epithelium of control group,but the expression of PAX8, PAX2, p53 and RAS were strongly positive brown in secrete cells of normal fallopian tube epithelium.(2)p53 and RAS expression of fallopian tube epithelium in the epithelial ovarian cancer group were significantly higher than those in the non-epithelial ovarian cancer groups(P<0.05),but the expression of PAX8 and PAX2 in fallopian tube and the expression of PAX8,PAX2,p53 and RAS in ovarian tissue was not statistically significant in the groups(P>0.05).PAX8,PAX2 and p53 expression of the ovarian in HGSC group were significantly higher than those in LGSC group(P<0.05),while the expression of RAS was lower in the ovarian of the high-grade group (P<0.05), while the expression of PAX8, PAX2, p53 and RAS in fallopian tube was not statistically significant in the groups(P>0.05).(3)There was a significantly positive correlation between fallopian tube and the corresponding ovary of HGSC in PAX8 and PAX2 expression(r=0.422, P=0.045; r=0.693, P=0.000), but not correlation in p53 and RAS expression (r=0.058, P=0.793; r=-0.190,P=0.384).(4)Univariate survival analysis showed that the progression free survival time in patients with ovarian serous cancer group was significantly correlated with the protein expression of PAX8, PAX2 and RAS(P<0.05),but there were not correlated with age,surgical staging,cell differentiation,lymph node metastasis and preoperative chemotherapy and p53 protein expression (P>0.05). The total survival time in patients with ovarian serous cancer group was significantly correlated with the protein expression of PAX8 (P<0.05),but there were not correlated with age,surgical staging,cell differentiation,lymph node metastasis and preoperative chemotherapy and the protein expression of PAX2, RAS and p53 (P>0.05). Conclusions PAX8, PAX2, p53, RAS are of great significance for the study of origin of HGSC. HGSC may be derived from fallopian tube, but further investigation would be necessary to confirm this. PAX8, PAX2, p53, RAS could be expected to be used as predictors of survival prognosis in patients with ovarian serous cancer.

Síndrome cardio-facio-cutáneo: reporte de caso / Cardio-facio-cutaneous syndrome: a case report

Resumen: el síndrome cardio-facio-cutáneo es una entidad clínica y genéticamente heterogénea, perteneciente a un grupo de síndromes conocidos como RASopatías.Este trastorno es de baja prevalencia, con alrededor de 200 a 300 casos en el mundo, e incluye entre sus manifestaciones clínicas rasgos faciales dismórficos, defectos cardíacos y alteraciones cutáneas. Los hallazgos fenotípicos del síndrome cardio-facio-cutáneo que se comparten con otros síndromes y la ausencia de criteriosdiagnósticos o signos patognomónicos lo convierten en un reto diagnóstico. En este manuscrito se presenta un caso confirmado de síndrome cardio-facio-cutáneo por estudios de genética molecular en una paciente de siete años de edad, mediante el cual se exponen las principales características de esta condición.

Abstract: The cardio-facio-cutaneous syndrome is a clinically and genetically heterogeneous disorder, belonging to a group of syndromes known as RASopathies. This condition has a low prevalence, with around of 200 to 300 cases in the world, and includes dysmorphic facial features, heart defects, and skin abnormalities among its clinical manifestations. The phenotypic findings of cardio-facio-cutane1ous syndrome that are shares with other syndromes and the absence of diagnostic criteria or pathognomonic signs make it a diagnostic challenge. Here its present a confirmed case of cardio-facio-cutaneous syndrome by molecular genetic studies in one seven years old patient, through which are exposed the main characteristics of this condition.

Result of 2014 external quality assessment for KRAS mutation testing / 中华检验医学杂志

Objective To evaluate the performance of KRAS gene mutation detection in 2014 external quality assessment ( EQA ) program and discuss the problems in clinical laboratories .Methods The sample panel of 2014 EQA program contained 5 artificial formalin-fixed, paraffin-embedded ( FFPE) samples.The participating laboratories were asked to report their results before the deadline .The scores of EQA and the rate of overall coincidence , false positive and false negative were calculated .Results The EQA program for KRAS testing was set twice a year .In 2014, 58 and 57 valid lab results were submitted respectively.About 79.31%(46/58)and 94.73%(54/57) of the laboratories were correct for all samples. The coincidence rate of positive samples were 93.53% ( 217/232 ) and 96.49% ( 165/171 ) . The coincidence rate for negative ones were 100%(58/58) and 98.25% (112/114).The false-negative ratio was 1.29%( 3/232 ) and 0%.The false-positive ratio was 4.14% ( 12/290 ) and 3.15% ( 9/285 ) . Conclusions The results of 2014 EQA for KRAS gene mutation testing suggested that the performance of laboratories had been improved significantly , however , the false-negative and false-positive results had always been the major problems affecting the accuracy of KRAS mutations testing .Laboratories needed to standardize the testing process and manufacturers should optimize the reagents and the way of interpretation , to guarantee the performance of KRAS gene mutation detection .

Progress in the detection of Kras, EGFR, BRAF mutations and its clinical application / 中华检验医学杂志

Since a number of genes have been discovered,which are associated with the effect of drug therapy,the development of personalized medicine is enhanced.Meanwhile,personalized test played an important role in choosing proper medicine.Herein,the status of detection of V-ki-ras2 Kirsten rat sarcoma viral oncogene homolog(Kras),epidermal growth factor receptor(EGFR),B-Raf and v-Raf murine sarcoma viral oncogene homolog B1(BRAF) mutation is concluded,moreover,the association between the mutation test and the effect of cancer therapy is summarized.

Studies on the expressions and clinical significance of miRNAlet 7 and Ras in non-small cell lung cancer / 中国医师杂志

Objective To study the expressions of miRNAlet7 and Ras in non-small cell lung cancer ( NSCLC),and their correlations with clinicopathological features and survival time.Methods In-situ hybridization was used to detect the expression of let7,and SP immunohistochemistry to measure HMGA2 in 68 NSCLC cases ( group A) and 20 cases with normal lungs ( group B).Results The positive rate of let7 in group A was lower than that in group B (39.7％ vs 63.2％ ) ( P ＜0.05).The positive rate of Ras in group A was higher than that in group B (66.2％ vs 25.0％ ) ( P ＜0.01 ).The positive rate of let7 was not related to the age,gender,histological type,cell differentiation,and clinical stages of cancer patients(P ＞0.05).The positive rate of Ras was related to smoking,sex,and histological type of cancer( P ＜0.01 ),and was not related to cell differentiation,lymphatic metastasis,and clinical stages of cancer ( P ＞0.05).There was an obvious negative correlation between let7 and Ras( r =-0.627,P ＜0.01 ).The 2-year survival rate of let7-positive group was higher than that of the let 7-negative group ( x2 =4.84,P ＜0.05).No statistically significant difference was found between Ras-positive and-negative groups ( P ＞0.05 ).Conclusions The lower level of let7 expression,and high level of Ras expression have much to do with the carcinogenesis of NSCLC.The level of let7-positive expression is closely related to prognosis; while Ras may act in cooperativity in the occurrence and development of NSCLC.

Molecular Docking and Simulation studies of Farnesyl Trasnferase with the potential inhibitor Theflavin.

Ras proteins play crucial roles in cell growth regulation, signal transduction and proliferation. Abnormal Ras proteins caused by genetic mutations are more common in human cancers. Activation of Ras occurs by enzymatic attachment of Farnesyl moiety by Farnesyl transferase. It has been proven that Ftase is a potential anticancer protein target. In this study we perform a virtual screening using polyphenol derivative from various sources against Ftase and establish ‘Theaflavin’ an antioxidant polyphenol extracted from Camellia sinensis (tea), as a potential inhibitor. This conclusion is further supported by comparative studies of molecular docking and 10 nS molecular dynamics simulation with ‘Tipifarnib’ (Zarnestra) a preclinical Ftase inhibitor.

The construction of pcDNA 3.1 RASSF1 eukaryotic vector and the influence of RASSF1 on apoptosis of hepatocarcinoma cell line / 中国医师杂志

Objective To construct pcDNA3.1 RASSF1 eukaryotic vector and observe the influence of RASSF1 on the apoptosis of hepatocarcinoma cell line HepG2. Methods RASSF1 gene was amplifled from human RASSF1 cDNA by polymerase chain reaction (PCR) and cloned into pcDNA3.1. The recombinant plasmid pcDNA3. 1 RASSF1 was transfected into hepatocarcinoma HepG2 cell line. The expression of RASSF1 was examined by Western blot. The influence of RASSF1 on the cell apoptosis was measured by Annexin V/PI assay. Results DNA enzyme digestion and sequencing results showed that recombinant plasmid pcDNA3. 1-RASSF1 was successfully constructed. RASSF1 protein was overexpressed in HepG2 cell line transfected with pcDNA3. 1-RASSF1 plasmid. The apoptosis rate of blank, pcDNA3. 1 and pcDNA3. 1-RASSF1 group was (5.8 ±0.42)%, (7.48 ±0.68)% and (35. 1 ±3. 15)%, respectively.Conclusion The pcDNA3. 1- RASSF1 eukaryotic vector was successfully constructed, RASSF1 protein overexpression could induce apoptosis in HepG2 cell line.

Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma

INTRODUCTION: Epidermal growth factor is a single-chain polypeptide consisting of 53 amino acids with potent mitogenic activity that stimulates the proliferation of a range of normal and neoplastic cells through an interaction with its specific receptor (epidermal growth factor receptor, EGFR). This interaction plays a key role in tumor progression including the induction of tumor cell proliferation. An increased EGFR copy number have been associated with a favorable response to EGFR tyrosine kinase inhibitors therapy. In contrast, K-ras mutations tend to predict a poor response to such therapy. The aim of this study was to determine the correlation between the clinicopathological factors and the up-regulation of EGFR expression and Kras mutations in oral squamous cell carcinoma. MATERIALS AND METHODS: This study examined the immunohistochemical staining of EGFR, K-ras mutation detection with peptide nucleic acid (PNA)-based real-time polymerase chain reaction (PCR) clamping in 20 specimens from 20 patients with oral squamous cell carcinoma. RESULTS: 1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, a high level of EGFR staining was observed. The correlation between immunohistochemical EGFR expression and histological differentiation, as well as the tumor size of the specimens was significant (Pearson correlation analysis, significance [r] >0.5, P<0.05). 2. In PNA-based real-time PCR clamping analysis, a K-ras mutation was not detected in all specimens. CONCLUSION: These findings suggest that the up-regulation of the EGFR may play a role in the progression and invasion of oral squamous cell carcinoma that is, independent of a K-ras mutation.

The effect of Ras protein on the delayed myocardial protection of Lovastatin preconditioning in the ischemia and reperfused rat hearts / 中国医师杂志

Objective To investigate the effect of Ras protein on the delayed myocardial protection of Lovastatin preconditioning in the ischemic and reperfused rat hearts.Methods 24 Sprague-Dawley male rats were randomly and evenly divided into three groups,model control group,N group and Lovastatin group.Lovastatin group was treated with 15mg Lovastatin per a kilogram once a day,N group treated with the same dose of Lovastatin with Lovastatin group and L-NAME 30mg per a kilogram once a day.Rat models of I/R were established with coronary occlusion 30 minutes and reperfusion 30 minutes of the left anterior descending artery after two-week's administration.The expression of Ras-GTPase protein in the heart tissue was determined and myocardial apoptosis was detected in all groups.Results The number of TUNEL positive myocardial nuclei in L group was remarkably reduced,compared with C group(t=2.32,P＜0.05).Ras-GTPase protein in the heart tissue was significantly inhibited in the L and N group,compared with C group(t=2.25,P＜0.05).Conclusion Lovastatin may have the delayed protective effect on the ischemie and reperfused rat heart,which effect may be correlated with inhibition of RasGTPase protein during the period of ischemia and reperfusion.Its protective effect was not correlated with NO.

Effects of sulphated heparin on proliferation and apoptosis of human hepatocellular carcinoma cell line (HepG2) / 中国病理生理杂志

AIM: To explore the effect and the mechanism of sulphated heparin on the proliferation and the apoptosis of human hepatocellular carcinoma cells. METHODS: The human hepatocellular carcinoma cell line (HepG-2) was used to identify the expression of ras gene protein and to study the effect of sulphated heparin on proliferation and the apoptosis in vitro . RESULTS: The sulphated heparin downregulated the ras protein expression and inhibited the cell growth in HepG2 cells. In the presence of sulphated heparin, the apoptosis rate of HepG2 increased. CONCLUSION:The data suggest that the effects of sulphated heparin on the proliferation and the apoptosis of the human hepatocellular carcinoma cell are correlated with the signaling transduction mediated by ras gene protein.