[Effect of anti-HPV16 E6-ribozyme on the sensitivity of cervical carcinoma cell line to chemotherapy].
OBJECTIVE
To study the role of anti-HPV16E6-ribozyme in modulating the sensitivity of cervical carcinoma cell line to chemotherapy.


METHODS
By way of lipofectin transfection, anti-HPV16E6-ribozyme antibody and empty eukaryotic expression plasmids were respectively transfected into CaSKi cells that were designated as CaSKi-R and CaSKi-P cells accordingly. Ribozyme expression in the transfected cells was subsequently observed with RNA dot blot, and the amount of E6 mRNA expression detected by Northern blotting. Cell count was performed for determining the growth rate of the non-transfected and transfected CaSKi cells and colony formation test was employed to examine the sensitivity of the cells to chemotherapy. PI/Annexin V staining was conducted to determine the apoptosis rates of each cell line in response to chemotherapy.


RESULTS
As shown by dot blot analysis, stable expression of anti-HPV16E6-ribozyme was achieved in CaSKi-R cells, in which lower E6 mRNA expression level was observed by means of Northern blotting in comparison with those in CaSKi-P and CaSKi cells. Significant slow-down of the growth rate occurred in CaSKi-R cells as compared with the growth rate of the other 2 cell lines, but no differences in relative cloning efficiency and apoptosis rates of the 3 cell lines were observed in response to taxol treatment (P>0.05), while cis-platinum treatment produced comparatively deceased relative cloning efficiency and increased apoptosis rate in CaSKi-R (P<0.01).


CONCLUSIONS
Transfection by anti-HPVE6-ribozyme may inhibit the growth of CaSKi cells and increase their sensitivity to cis-platinum but not to taxol.
Rao
Zhi-Guo
ZG
Oncology Center, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
Zhang
Ji-Ren
JR
Zheng
Yan-Fang
YF
Zhou
Hui
H
Qu
Liang-Gu
LG
D000906:Antibodies / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D004347:Drug Interactions; D004354:Drug Screening Assays, Antitumor; D005260:Female; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D017239:Paclitaxel / Q000494:pharmacology; D012097:Repressor Proteins; D012270:Ribosomes / Q000276:immunology* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000473:pathology
D000906:Antibodies; D000970:Antineoplastic Agents; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins; D017239:Paclitaxel; D002945:Cisplatin