PiggyBac transposon system with polymeric gene carrier transfected into human T cells.

CAR-T cell-based immunotherapy has shown great promise in clinical trials for the treatment of hematological malignancies. The majority of these trials utilize retroviral and lentiviral vectors to introduce CAR transgene. In spite of its satisfactory efficiency, the concerns about the potential carcinogenicity and complicated synthesis procedure restrict widespread clinical applications of viral vectors. Recent studies show that transposon-based gene transfer is a safer and simpler non-viral approach for stable transgene expression. Here, we developed an in house made polymeric nanomicelles carrier for piggyBac (PB) transposon delivery to primary T lymphocytes. The properties, transfection efficiency and toxicity of this carrier was analyzed. Results indicated that nanomicelles produced in our study were stable and reduction-sensitive. These micelles can completely condense DNA and mediate transfection with efficiency of average 30.2% with high cell viability (> 80%). Furthermore, incorporating piggyBac transposase elements into polyplexes promoted persistent expression of the transgene (up to 55%). At the end of culture, CAR-T cells mainly exhibited memory phenotype and consisted of CD3+CD8+ T cells. The cytotoxicity of these CAR-T cells was average 17% at 20:1 ratio. In conclusion, polymeric nanomicelles provide a flexible and safe method for gene delivery to T lymphocytes.

Effect of Wubeizi ointment aqueous solution on the expression of type I and III procollagen genes in keloid fibroblasts.

We evaluated the effect of Wubeizi (WBZ) ointment on keloids. Keloid-derived fibroblast primary cultures were used to evaluate the effect of the different concentration of WBZ ointment on the expression of type I and III procollagen in keloid fibroblast primary cultures using dot blot assay. Type I and II precollagen cDNA probes labeled with non-radioactive digoxin were used for dot blot. Cell cultures were divided into 4 groups: The large dose group received 1 g/ml of WBZ, middle dose, and small dose groups received 0.5 and 0.25 g/ml of WBZ, respectively. The control group received serum-free medium without WBZ. Our results showed that type I and III procollagen mRNA expression was reduced significantly in the large dose and middle dose groups compared to the control group. Type I and III procollagen mRNA expression level in the small dose group had no statistically significant difference with the control group. However, the difference between the large dose group and the small dose group was statistically significant. We concluded that WBZ ointment aqueous solution restricted keloid fibroblast proliferation by downregulating the expression of type I and III procollagen and therefore reducing collagen deposition in keloid tissue.

Effects of resveratrol on the proliferation, apoptosis and telomerase ability of human A431 epidermoid carcinoma cells.

The aim of the present study was to investigate the effect of resveratrol on cell apoptosis, ability of telomerase and the human telomerase reverse transcriptase (hTERT) protein expression in human A431 epidermoid carcinoma cells. A431 cells were treated with different concentrations of resveratrol, and the cell appearance was then observed under a microscope. In addition, the cell proliferation was examined using an MTT assay, and the ability of telomerase was detected using telomeric repeat amplification protocol-polymerase chain reaction-ELISA. Resveratrol significantly inhibited the ability of telomerase and decreased the expression of hTERT protein in a concentration-dependent manner. In conclusion, resveratrol is capable of downregulating the expression of hTERT protein and inhibits the ability of telomerase of A431, which is an important mechanism of action of resveratrol with regard to inhibition of A431 cell proliferation.

The effects of Wubeizi ointment on the proliferation of keloid-derived fibroblasts.

UNLABELLED: To evaluate the effectiveness of the Wubeizi (WBZ) ointment on keloid-derived fibroblasts. The primary cells of the keloid-derived fibroblasts were cultured and the effectiveness of the WBZ ointment at different concentrations was examined by MTT colorimetric methods on keloid-derived fibroblasts. The WBZ ointment showed inhibitory effects on proliferating the keloid-derived fibroblasts (P < 0.01)in a time- and dose-dependent manner. The proportion of cells in S stage was significantly higher in each of the WBZ ointment group than in the control group (P<0.01), and the proportion of G2 + M stage cells was significantly lower than that of control group, which was statistically significant (P < 0.01).The inhibitory effects of the S and G2 + M stage increased with higher drug concentrations (P < 0.05). CONCLUSION: The WBZ ointment can inhibit the proliferation of the keloid-derived fibroblasts in a time- and dose- dependent manner.