Abstract | BACKGROUND: METHODS: The concentration of SPARC was measured with an enzyme-linked immunosorbent assay (ELISA); distribution and expression levels were measured with in situ hybridization, immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and western blot assays. Apoptosis was assessed by morphological observation and fluorescence-activated cell sorting (FACS) apoptosis index (AI) analysis. Cell cycle phase was examined by FACS analysis. Cell proliferation was studied using bromodeoxyuridine ( BrdU) incorporation ELISA. RESULTS: The SPARC level in the renal cyst fluid of patients with ADPKD was greater than that in patients with simple renal cyst (SRC), and also greater than that found in the plasma and urine of patients with either ADPKD or SRC and normal subjects. SPARC mRNA and protein levels in polycystic renal tissue were greater than that in normal renal tissue. Additionally, SPARC could inhibit cyst-lining epithelial cell proliferation, bring about cell cycle arrest in the G0/G1 phase and induce apoptosis in vitro. SPARC treatment resulted in decreased mRNA levels of PCNA ( proliferating cell nuclear antigen), MCM2 (minichromosome maintenance protein 2), ClnD1 and Bcl-2, but an increased mRNA level of p21(Waf1) in cyst-lining epithelial cells. CONCLUSION: Our findings suggest that the increased SPARC expression in ADPKD renal tissue may provide negative feedback in ADPKD patients.
|
Authors | Wenjing Wang, Changlin Mei, Bing Tang, Haidan Zhao, Chenggang Xu, Zhanyuan Li, Xuefei Shen, Wencheng Fu, Bing Dai |
Journal | Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association
(Nephrol Dial Transplant)
Vol. 21
Issue 5
Pg. 1278-88
(May 2006)
ISSN: 0931-0509 [Print] England |
PMID | 16421164
(Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- DNA, Complementary
- Osteonectin
|
Topics |
- Adolescent
- Adult
- Aged
- Apoptosis
(genetics)
- Base Sequence
- Blotting, Western
- Case-Control Studies
- Cell Proliferation
- Cells, Cultured
- DNA, Complementary
(analysis)
- Enzyme-Linked Immunosorbent Assay
- Epithelial Cells
(cytology)
- Female
- Gene Expression Regulation
- Humans
- Immunohistochemistry
- In Situ Hybridization
- In Vitro Techniques
- Male
- Middle Aged
- Molecular Sequence Data
- Osteonectin
(genetics)
- Polycystic Kidney, Autosomal Dominant
(genetics, pathology)
- Probability
- Reference Values
- Reverse Transcriptase Polymerase Chain Reaction
- Sampling Studies
|