Adolescent ; Adult ; DNA ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 ; Male ; Mutation ; Parkinsonian Disorders ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics

OBJECTIVE: To screen for pathogenic variants in the coding regions of STK11 gene among Chinese patients with Peutz-Jeghers syndrome (PJS). METHODS: Peripheral blood samples were collected from 64 patients. The coding regions of the STK11 gene were detected by PCR and Sanger sequencing. RESULTS: Fourty-eight patients were found to harbor STK11 gene variants, which included 39 types of variants consisting of missense, nonsense, insertional, deletional and splice site variants. Among 64 PJS patients, the detection rate of point variants was 75.00% (48/64), of which missense variants accounted for 29.17% (14/48), nonsense variants accounted for 29.17%(14/48), insertion variants accounted for 2.08% (1/48), deletional variants accounted for 10.42% (5/48), and splice site variants accounted for 29.17% (14/48). The detection rates of sporadic cases and those with a family history were 71.8% (28/39) and 80.0% (20/25), respectively. Two variants (c.250A>T, c.580G>A) occurred in 3 PJS probands. Thirteen variants were unreported previously and were considered to be pathogenic. CONCLUSION The detection rate of variants among Chinese PJS patients is similar to that of other countries. A number of novel common variant sites were discovered, which enriched the spectrum of PJS-related variants.
Asian Continental Ancestry Group ; China ; DNA Mutational Analysis ; Humans ; Peutz-Jeghers Syndrome ; genetics ; Protein-Serine-Threonine Kinases ; genetics

OBJECTIVETo detect protein expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2) in human hepatocellular carcinoma and adjacent non-neoplastic liver.

METHODSIn 16 surgically resected hepatocellular carcinoma and para-carcinoma tissues, Western blotting was used to detect expression of ERK(1), ERK(2), JNK(1), p38 and MEK(1), MEK(2).

RESULTSIn all cases, ERK(1), ERK(2), p38 expression in hepatocellular carcinoma was significantly higher than that in para-carcinoma: integral optic density (IOD) of ERK(1) was 300 +/- 98 in carcinoma and 98 +/- 48 in para-carcinoma tissues (t = 2.519, P < 0.01); IOD of ERK(2) was 587 +/- 83 in carcinoma and 232 +/- 96 in para-carcinoma tissues (t = 2.745, P < 0.01); IOD of p38 was 270 +/- 85 in carcinoma and 107 +/- 88 in para-carcinoma tissues (t = 2.491, P < 0.01). JNK(1) expression in hepatocellular carcinoma was significantly lower than that in para-carcinoma; IOD of JNK(1) was 111 +/- 93 in carcinoma and 292 +/- 109 in para-carcinoma tissues (t = 2.473, P < 0.01). Protein levels of MEK(1) and MEK(2) in carcinoma were significantly higher than in para-carcinoma. IOD of MEK(1) was 1 418 +/- 244 in carcinoma and 806 +/- 90 in para-carcinoma tissues (t = 2.546, P < 0.01). IOD of MEK(2) was 1 041 +/- 122 in carcinoma and 468 +/- 40 in para-carcinoma tissues (t = 2.861, P < 0.01).

CONCLUSIONSERK(1), ERK(2), MEK(1) and MEK(2) in the signal transduction pathway for cell proliferation are significantly overexpressed and the expression of JNK(1) is lower in hepatocellular carcinoma. Their unbalance is one of the important reasons for the over growth and infinite proliferation of the hepatocellular carcinoma cell. The p38 and JNK(1) may be activated by different pathway.

Adult ; Aged ; Carcinoma, Hepatocellular ; enzymology ; Enzyme Activation ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases ; Liver Neoplasms ; enzymology ; MAP Kinase Kinase 1 ; Male ; Middle Aged ; Mitogen-Activated Protein Kinase Kinases ; analysis ; Mitogen-Activated Protein Kinases ; metabolism ; Protein-Serine-Threonine Kinases ; analysis

The ras, is a G-like protein that controls the mitogen-activated protein kinase (MAPK) pathway involved in control and differentiation of cell growth. MAPK is a key component of its signaling pathway and the aberrant activation may play an important role in the transformation process. To better understand roles of ras in the activation of MAPKs, we have established ras transformed NIH3T3 fibroblast cell line, and analyzed the MAPK module. The ras transformed cells formed numerous spikes at the edges of cells and showed loss of contact inhibition. The levels of ERK1/2 MAPKs as revealed by Western blot analysis were not significantly different between ras transformed and non-transformed cells. However, phosphorylation of ERK MAPKs and the level of MEK were significantly increased although the heavily expressed level of Raf-1, an upstream component of MAPK pathway was unchanged in ras transformed NIH3T3 cells. The sedimentation profile of the MAPK module kinases in a glycerol gradient showed the presence of a rather homogeneous species of multimeric forms of ERK1/2 and MEK as indicated by the narrow distribution peak areas. The broad sedimentation profile of the Raf-1 in a glycerol gradient may suggest possible heterologous protein complexes but the identification of interacting molecules still remains to be identified in order to understand the organization of the MAPK signal transduction pathway.
3T3 Cells ; Animal ; Cell Transformation, Neoplastic* ; Genes, ras* ; Mice ; Mitogen-Activated Protein Kinase Kinases/analysis ; Mitogen-Activated Protein Kinases/analysis ; Phosphorylation ; Protein-Serine-Threonine Kinases/analysis* ; Proto-Oncogene Proteins c-raf/analysis ; p42 MAP Kinase/analysis

OBJECTIVETo investigate STK15 gene abnormality and centrosomal amplification in laryngeal carcinoma.

METHODSSTK15 gene mRNA expressional level was tested in 62 cases of laryngeal squamous cell carcinoma and laryngeal squamous cell carcinoma cell line Hep-2 by reverse transcription-polymerase chain reaction(RT-PCR); the mutation of STK15 gene exon 6 and exon 7 in the same tissues and cells was detected by PCR-single strand conformation polymorphism. Immunofluorescent antibodies were used to test centrosomal amplification in Hep-2 cell line as an example.

RESULTSSTK15 gene overexpressed in 39 cases of laryngeal carcinoma (63%) and Hep-2 cell line. No mutation was found in exon 6 and exon 7 of STK15 gene in the above tissues and cells. Centrosomal amplification was apparent in Hep-2 cell line. The number of centrosome in a single cell changed from 1 to 7, and Hep-2 cells with amplified centrosomes (more than 2 in one cell) were 11%-23%.

CONCLUSIONSTK15 gene overexpression and centrosomal amplification were first found in human laryngeal squamous cell carcinoma, which indicated that STK15 gene overexpression leading to centrosomal amplification might occur in the early stage of human laryngeal carcinogenesis and be one of the key mechanisms for the occurrence of laryngeal carcinoma.

Aurora Kinase A ; Aurora Kinases ; Centrosome ; pathology ; Exons ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Mutation ; Protein-Serine-Threonine Kinases ; genetics ; RNA, Messenger ; analysis

OBJECTIVETo investigate the expressions of cyclin E, cyclin dependent kinase 2 (CDK-2) and cyclin-dependent kinase inhibitor p57(KIP2) in human gastric cancer, and to evaluate the relationships between protein levels and clinicopathological parameters.

METHODSWestern blot was used to measure the expressions of cyclin E, CDK-2 and p57(KIP2) proteins in the surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 36 patients.

RESULTSCyclin E and CDK-2 protein levels were higher in gastric cancer tissues in comparison with normal tissues (P < 0.05). Overexpression of cyclin E was correlated with lymph node involvement, poor histological grade and serosa invasion (P < 0.05). Overexpression of CDK-2 was correlated with lymph nodes involvement (P < 0.05). No statistically significant difference between cyclin E and CDK-2 expression was found when samples were stratified according to tumor size (P > 0.05). Expression of cyclin E and CDK-2 showed a positive linear correlation (r = 0.451, P = 0.01). Protein levels of p57(KIP2) were lower in gastric cancer tissues than in the normal mucosa (P < 0.05). Decreased expression of p57(KIP2) was correlated with lymph node involvement (P < 0.05). No statistically significant difference in p57(KIP2) expression was found when sample were stratified according to tumor size, histological grade or serosa invasion (P > 0.05). In metastatic lymph nodes, expression of cyclin E was increased and the expression of p57(KIP2) decreased.

CONCLUSIONOverexpressions of cyclin E, CDK-2 and downregulated expression of p57(KIP2) may play important roles in tumorigenesis and metastatic potential of gastric cancer.

Blotting, Western ; CDC2-CDC28 Kinases ; Cyclin E ; analysis ; physiology ; Cyclin-Dependent Kinase 2 ; Cyclin-Dependent Kinase Inhibitor p57 ; Cyclin-Dependent Kinases ; analysis ; physiology ; Humans ; Lymphatic Metastasis ; Nuclear Proteins ; analysis ; physiology ; Protein-Serine-Threonine Kinases ; analysis ; physiology ; Stomach Neoplasms ; chemistry ; pathology

OBJECTIVETo analyze the sequence of STK11 gene coding region in 20 patients with Peutz-Jeghers syndrome and identify the point mutations in STK11 gene associated with the occurrence of the disease.

METHODSBlood samples were collected from 20 inpatients with Peutz-Jeghers syndrome treated in our center between January 2009 and October 2010. The sequence of STK11 gene coding region was analyzed using PCR and DNA sequencing and compared with the normal sequence of STK11 gene.

RESULTSOf the 20 patients with Peutz-Jeghers syndrome, 14 showed STK11 gene mutations in the coding region, including 1 patient having two mutations and 13 patients with a single mutation site. In one case, sequence analysis of the STK11 gene identified a novel type of STK11 germline mutation, in which the cytosine (C)460 was substituted by guanine (G) in exon 3 to result in a new amino acid at codon 154. Four patients from 2 families were found to have a common mutation. The remaining 6 patients were not found to have mutations in STK11 gene coding region.

CONCLUSIONMutations of STK11 gene is a major cause of Peutz-Jeghers syndrome. The missense mutation of 460 C→G in exon 3 of STK11 gene is a novel mutation associated with Peutz-Jeghers syndrome.

Adult ; Asian Continental Ancestry Group ; genetics ; Codon ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Peutz-Jeghers Syndrome ; genetics ; Protein-Serine-Threonine Kinases ; genetics

OBJECTIVERecent studies have shown that integrin linked kinase (ILK) plays an important role in the pathogenesis and development of some kidney diseases. This study aimed to investigate the relationship between ILK and renal glomerular damage in children with Henoch schonlein purpura nephritis (HSPN).

METHODSOne hundred and eighty eight HSPN children (aged 3 to 17 years) were assigned to five groups according to the classification of the International Study of Kidney Disease in Children (ISKDC): grade < or = IIa (n = 62), grade IIb (n = 42), grade IIIa (n = 29), grade IIIb (n = 40) and grade > or = IV (n = 15). Fifteen children with basement membrane nephropathy served as the control group. ILK expression on glomeruli was ascertained by immunohistochemical staining. The relationships of ILK expression on glomeruli with glomerular histopathologic lesions and urinary protein excretions were examined.

RESULTSThe positive areas of ILK expression on glomeruli in the control, grade < or = IIa, grade IIb, grade IIIa, grade IIIb and grade > or = IV groups were (3.35 + or - 1.01)%, (4.88 + or - 1.13)%, (9.64 + or - 1.36)%, (11.27 + or - 1.68)%, (17.42 + or -3.0)% and (20.62 + or - 2.32%), respectively. There were significant differences in the ILK expression between groups (p<0.01). ILK expression on glomeruli increased with increased urinary protein excretions. There were significant differences in the ILK expression in children with different urinary protein excretions (p<0.01).

CONCLUSIONSILK might be involved in the process of renal glomerular histopathologic damage and the production of proteinuria in children with HSPN.

Child ; Child, Preschool ; Female ; Humans ; Immunohistochemistry ; Kidney Glomerulus ; pathology ; Male ; Nephritis ; enzymology ; pathology ; Protein-Serine-Threonine Kinases ; analysis ; Purpura, Schoenlein-Henoch ; enzymology ; pathology

OBJECTIVETo investigate the expression of cancer-related gene MPS-1 in gastric cancer and to evaluate its significance in clinical diagnosis and therapy.

METHODSThe mRNA expression of MPS-1 was determined by polymerase chain reaction after reverse transcription (RT-PCR) in cancer tissues and adjacent non-cancerous tissues from 42 cases with gastric cancer. The expression levels of MPS-1 in 6 gastric cancer cell lines (AGS, MKN-45, SGC 7901, KATO III, N-87 and SNU-1) were also determined by RT-PCR and Western blot.

RESULTSThe MPS-1 mRNA was expressed in all tissues and cell lines. The mRNA expression level of MPS-1 in cancer tissues were 1.37+/- 0.87, significantly higher than 0.99+/- 0.67 in adjacent normal gastric mucous tissues (P< 0.01). The expression of MPS-1 was correlated with TNM stage (P< 0.05), but not with age, gender, tumor size and differentiation. The expression level of MPS-1 mRNA in the primary lesions was hig her in the patients with TNM stages III, IV than those with TNM stages I, II. Meanwhile, RT-PCR and Western blot showed the same results that MPS-1 expression was higher in the six gastric cancer cell lines as compared with that in the normal gastric cell line GES-1.

CONCLUSIONThe high expression of MPS-1 in gastric cancer indicates that MPS-1 might play an important role in gastric carcinogenesis,which may provide a new target in immunotherapy for gastric cancer.

Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Female ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Protein-Serine-Threonine Kinases ; genetics ; Protein-Tyrosine Kinases ; RNA, Neoplasm ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; metabolism ; pathology

Angiotensin II ; pharmacology ; Blotting, Western ; Cells, Cultured ; Glomerular Mesangium ; cytology ; drug effects ; metabolism ; Humans ; I-kappa B Kinase ; NF-kappa B ; analysis ; Peptide Fragments ; pharmacology ; Protein-Serine-Threonine Kinases ; analysis