1.Telomerase activity and homozygous deletions of the p16 gene in liver metastases of colorectal carcinoma
JP MA ; WH ZHAN ; JS PENG ; YL HE ; ZQ ZHENG ; SR CAI ; L WANG ; JP WANG
Chinese Medical Journal 2001;114(10):1068-1072
Objective To study the interaction between telomerase activity and abnormalities of the p16 gene in liver metastases of colorectal carcinoma. Methods Telomerase activity was detected by a non-isotopic PCR-based telomeric repeat amplification protocol (TRAP) assay, and homozygous deletions of the p16 gene were detected by a semiquantitative multiplex polymerase chain reaction in tissue samples from 24 liver metastases of colorectal carcinoma and 5 primary colorectal carcinomas. Results Telomerase activity was observed in 19 (79.2%) of 24 liver metastases of colorectal carcinoma. Telomerase activity was also observed in all 5 primary colorectal carcinomas and in 3 of their liver metastatic samples. The incidence of telomerase activity in liver metastases of colorectal carcinoma was not significantly correlated to tumor diameter, number of tumors, cirrhosis, and HBsAg. Homozygous deletions of the p16 gene were found in 9 of 24 (37.5%) liver metastases of colorectal carcinoma. Homozygous deletions of the p16 gene were observed in 2 of the 5 primary colorectal carcinomas and in 1 of the matching liver metastatic cancers. There was a correlation between telomerase activity and homozygous deletions of the p16 gene. Conclusions There is a correlation between telomerase activity and homozygous deletions of the p16 gene in liver metastases of colorectal carcinoma, suggesting its crucial role in liver metastases. However, telomerase activation and homozygous deletions of the p16 gene might not be the initiating event in liver metastases of colorectal carcinoma.
2.Immune privilege induced by cotransplantation of islet and allogeneic testicular cells
P LAN ; WH ZHAN ; JP WANG ; XJ WU ; LN YAN ; LJ XIAO
Chinese Medical Journal 2001;114(10):1026-1029
Objective To induce islet allograft long-term survival through cotransplantation of islet cells with sertoli cells. Methods Testicular sertoli cells were prepared by digestion with collagenase, trypsin and DNase, and were cultured for 48 hours. Collagenase digested and Ficoll purified donor (Wistar rat) islets were cotransplanted with allogeneic sertoli cells in the absence of systemic immunosuppression. Terminal leoxynucleotidyl transferase-mediated X-dUTP nick-end labeling (TUNEL) was used to label apoptosis of lymphocytes surrounding the islet graft. Results Cotransplantation of islets and 1 × 107 sertoli cells reversed the diabetic state for more than 60 days in 100% (6/6) of the chemically diabetic Sprague Dawley rats. Grafts consisting of islets alone or islets plus 1 × 105 sertoli cells survived only for 5 - 6 days. Apoptosis of lymphocytes surrounding the islets was quite clear. Conclusion Cotransplantation of islets with FasL+ sertoli cells induces local immune privilege and allows long-term graft survival without systemic immunosuppression.
3.Experimental Study of Cell Migration and Functional Differentiation of Transplanted Neural Stem Cells Co-labeled with Superparamagnetic Iron Oxide and Brdu in an Ischemic Rat Model
ZHU WEN-ZHEN ; LI XIANG ; QI JIAN-PIN ; TANG ZHOU-PING ; WANG WH ; WEI LI ; LEI HAO
Biomedical and Environmental Sciences 2008;(5):420-424
Objective To explore the migration of transplanted neural stem cells co-labeled with superparamagnetic iron oxide (SPIO) and bromodeoxyuridine (Brdu) using the 4.7T MR system and to study the cell differentiation with immuno-histochemical method in ischemic rats. Methods Rat neural stem cells (NSCs) co-labelled with SPIO mediated by poly-L-lysine and romodeoxyuridine (BrdU) were transplanted into the unaffected side of rat brain with middle cerebral artery occlusion (MCAO). At weeks 1, 2, 3, 4, 5, and 6 after MCAO, migration of the labelled cells was monitored by MRI. At week 6, the rats were killed and their brain tissue was cut according to the migration site of transplanted cells indicated by MRI and subjected to Prussian blue staining and immunohistochemical staining to observe the migration and differentiation of the transplanted NSCs. Results Three weeks after transplantation, the linear hypointensity area derived from the migration of labelled NSCs was observed by MRI in the corpus callosum adjacent to the injection site. Six weeks after the transplantation, the linear hypointensity area was moved toward the midline along the corpus callosum. MRI findings were confirmed by Prussian blue staining and immunohistochemical staining of the specimen at week 6 after the transplantation. Flourescence co-labelled immunohistochemical methods demonstrated that the transplanted NSCs could differentiate into astrocytes and neurons. Conclusion MRI can monitor the migration of SPIO-labelled NSCs after transplantation in a dynamical and non-invasive manner. NSCs transplanted into ischemic rats can differentiate into astrocytes and neurons during the process of migration.