OBJECTIVETo explore the potential molecular mechanisms for Bushen Tiaojing Recipe (BTR) improving the endocrine function of ovarian granular cells by observing the effect of BTR containing serum on follicle stimulating hormone/cyclic adenosine monophosphate-protein kinase A (FSH/ cAMP-PKA) pathway in in vitro cultured human ovarian granular cells.

METHODSThe primary ovarian granular cells collected from in vitro fertilization-embryo transfer patients were cultured for 24 h. The human and rat serum containing different concentrations of BTR (low, medium, high dose), and their normal serums were co-incubated with ovarian granular cells for 48 h respectively, and then they were divided into the low, medium, high dose BTR groups and the control group. The levels of estradiol (E2), progesterone (P), and cyclic adenosine monophosphate (cAMP) in the culture medium were measured by radioimmunoassay. The protein expression of FSHR in ovarian granular cells was detected by Western Blot. The mRNA expression of follicle stimulating hormone receptor (FSHR) and P450 aromatase (P450arom) in ovarian granular cells were detected by Real-time PCR.

RESULTSIn human BTR containing serum groups: Compared with control group, the levels of E2 and cAMP in the culture medium were higher (both P < 0.05) in the medium and high dose BTR groups; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells increased (all P < 0.01), the mRNA expressions of P450arom in ovarian granular cells were higher (P < 0.05, P< 0.01) in the medium and high dose BTR groups. In rat BTR containing serum groups: Compared with the control group, the levels of E2 in the culture medium were higher (all P < 0.01), cAMP in the culture medium were higher (P < 0.05, P < 0.01) in the medium and high dose BTR group; the levels of P in the culture medium decreased in the medium dose BTR group (P < 0.01). The protein and mRNA expression of FSHR in ovarian granular cells were higher (all P < 0.01), the mRNA expression of P450arom in ovarian granular cells increased in the medium and high dose BTR groups (P < 0.05, P < 0.01).

CONCLUSIONBTR could possibly improve the endocrine function of ovarian granular cells by regulating main effector molecules FSHR, cAMP, P450arom, and E2 in FSH/cAMP-PKA pathway of ovarian granular cells.

Cells, Cultured ; Cyclic AMP-Dependent Protein Kinase Type I ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; metabolism ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Humans ; Serum ; chemistry ; Signal Transduction ; drug effects

Humans ; Mutation ; Cyclic AMP-Dependent Protein Kinase Catalytic Subunits/genetics*

No abstract available.
Alternative Splicing ; Biopsy ; Carney Complex/diagnosis/enzymology/*genetics/therapy ; Cyclic AMP-Dependent Protein Kinase RIalpha Subunit/*genetics ; DNA Mutational Analysis ; Genetic Predisposition to Disease ; Humans ; Magnetic Resonance Imaging ; Male ; *Mutation ; Pedigree ; Phenotype ; Protein Isoforms ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo explore the relationship between the overexpression of PKA RIalpha mRNA and cliniopathological parameters in lung cancer.

METHODSRT-PCR was used to detect the expression of PKA RIalpha mRNA in 54 cases with human lung cancer and matched normal tissues.

RESULTS(1) The expression of PKA RIalpha mRNA was significantly higher in cancer tissue (66.7%) than in normal tissues (20.4%) (P < 0.01). (2) The expression was significantly correlated with TNM stage (P < 0.01), being increased with TNM stage. (3) The expression was significantly higher in patients with positive lymph nodes than in those with negative lymph nodes (P < 0.01). (4) There were no significant associations of PKA RIalpha mRNA expression with histological type, differentiation grade or size of the tumor.

CONCLUSIONThis study indicates that the overexpression of PKA RIalpha mRNA may play an important role in the progression, metastasis and prognosis of lung cancer.

Adenocarcinoma ; metabolism ; secondary ; Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cyclic AMP-Dependent Protein Kinase RIalpha Subunit ; Cyclic AMP-Dependent Protein Kinases ; biosynthesis ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics

To evaluate the expression of cAMP-dependent protein kinase type I-alpha regulatory subunit (PRKAR1α) in non-small cell lung cancer (NSCLC) and its correlation with clinicopathological features.
 Methods: PRKAR1α expressions in 79 NSCLC patients and matched adjacent non-carcinoma tissues were analyzed by using qRT-PCR and immunohistochemistry.
 Results: The negative rates of PRKAR1α protein in NSCLC, lung squamous cell carcinoma (SCL) and lung adenocarcinoma (ACL) were 58.2%, 77.8%, 32.4%, respectively. Compared to the matched adjacent non-carcinoma tissues, there were significant differences in levels of PRKAR1α mRNA and protein in ACL (P<0.05), but not in SCL and overall NSCLC (P>0.05). The expression of PRKAR1α protein was positively correlated with histological type, TNM stage, and lymph node metastasis (P<0.05). Tumor size and histogenesis differentiation were not related to the decreased PRKAR1α (P>0.05).
 Conclusion: Low expression of PRKAR1α in ACL might be involved in the pathogenesis, which might serve as a novel diagnostic candidate.
Adenocarcinoma ; chemistry ; classification ; genetics ; Adenocarcinoma of Lung ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; chemistry ; genetics ; Carcinoma, Squamous Cell ; chemistry ; genetics ; Cyclic AMP-Dependent Protein Kinase RIalpha Subunit ; physiology ; Female ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Lung Neoplasms ; chemistry ; classification ; genetics ; Lymphatic Metastasis ; genetics ; Male ; Neoplasm Staging ; RNA, Messenger