Experimental Studies of Using Real-time Fluorescence Quantitative PCR and RT-PCR to Detect E6 and E7 Genes of Human Papillomavirus Type 16 in Cervical Carcinoma Cell Lines
Methods We selected 4 target mRNA sequences to the exon and interon of HPV-16 E6 mRNA, and synthesized dsDNA to construct the recombinant pSi-lencer1.0-U6 vector which expresses HPV-16 E6 short hairpin dsRNA. After transfected into HPV-16 DNA positive cervical cancer cell line CaSki, then observe the expression level of HPV-16 E6,E7 genes and their proteins as well as the inhibition effects of cell growth.
RESULTS: CTLs stimulated by the DC vaccine had high killing activity on autologous cervical cancer cells, with killing rates of 79.32%-89.27% which were obviously higher than that of lymphokine-activated killing cells (t≥2.89, P<0.05).
The ICCs induced by HSP 70 were shown significant cytotoxic activity (82.69±1.97%,62.11±1.61%)against HeLa cells and cervical carcinoma cells,and the cytotoxic activity (31.05±2.09%)of the ICC against HeLa cells could be blocked by anti HSP 70 antibody.
This article discusses the characteristics of the drug-resistant human cervical carcinoma cell line HeLa/MMC and investigates reversal effect of SDZ-PSC833(PSC833) and Verapamil (Ver) upon HeLa/MMC.
Cervical carcinoma cell lines containing from 1-3 to 600 copies of the HPV genome were used as positive controls and were scored positive in all assays by both Southern blotting and GP-PCR.
We investigated here the expression pattern of AP-2α, AP-2β, and AP-2γ, as well as that of the cellular AP-2 target gene c-erbB-2, in a series of cervical cancer cell lines.
We found that, with the exception of HPV-negative C33A cells, all investigated cervical cancer cell lines expressed all three AP-2 family members, although at varying levels.
Three of the five cell lines (ovarian cancer cell line BG-1, cervical cancer cell line ME-180, and melanoma cell line SK-MEL 28) were sensitive to both IFNs.Cervical cancer cell line CaSki was sensitive to IFN-α2b but resistant to IFN-γ.
Cervical cancer cells contain detectable levels of functional p53 protein despite of the expression of the HPV E6 protein, which can induce p53 degradation.
These results indicate that p53 mutations are rare events in recurrent cervical carcinomas, and that somatic mutations of p53 do not provide cervical cancer cells with a selective growth advantage for recurrence.
Conclusions: We were able to reprogramme the target specificity of recombinant Cl96 and na?ve CTLs resulting in efficient cytolysis of CD44v7/8-positive cervical cancer cells.
Activation of p53 in cervical carcinoma cells was accompanied by induction of p53-dependent CDKN1 (p21), inhibition of cell proliferation, and induction of apoptosis.
McCoy cells (mouse fibroblasts), HeLa 229 (derived from human cervical carcinoma cells) and BHK-21 cells (baby hamster kidney cells) are the cell types regularly used for the culture ofC.