ABSTRACT
BACKGROUND: Bone marrow mesenchymal stem cells are characterized by wide sources and low immunogenicity. Especially, these cells are easy to import and express foreign genes, and thus have obvious superiorities as anti-tumor gene therapy vectors. OBJECTIVE: To investigate the effects of interleukin-12 gene modified bone marrow mesenchymal stem cells on the proliferation, cell cycle and apoptosis of ovarian cancer cells. METHODS: Interleukin-12 recombinant adenovirus vector was used to transfect bone marrow mesenchymal stem cells. Then, the expression of interleukin-12 mRNA and protien in transfected bone marrow mesenchymal stem cells was detected by RT-PCR and western blot assay, respectively. The level of interleukin-12 in cell supernatant was determined by ELISA. The SKOV3 cells were co-cultured with the supernatant of bone marrow mesenchymal stem cells transfected with (transfection group) or without (control group) interleukin-12 recombinant adenovirus vector. The proliferation of SKOV3 cells was determined by MTT assay. Flow cytometry was used to detect the cell cycle and apoptosis of SKOV3 cells. RESULTS AND CONCLUSION: RT-PCR and western blot results showed that interleukin-12 mRNA and protein were expressed in transfected bone marrow mesenchymal stem cells, but not found in empty vector group and blank control group. ELISA results showed that the content of interleukin-12 in the supernatant of bone marrow mesenchymal stem cells was (68.78±12.35) μg/L in the interleukin-12 transfection group after 48 hours culture, and no interleukin-12 expression was detected in the empty vector group and the blank control group, Interleukin-12-transfected bone marrow mesenchymal stem cells significantly inhibited the proliferation of SKOV3 cells, and the proliferation inhibition rate was increased with time (P < 0.05). The proportion of G1-phased SKOV3 cells was higher in the transfection group than in the control group (P < 0.05), and the percentage of G2-phased SKOV3 cells was lower in the transfection group than in the control group (P < 0.05). The apoptosis rate of SKOV3 cells in the transfection group was higher than that in the control group (P < 0.05). Our experimental findings indicate that bone marrow mesenchymal stem cells transfected with interleukin-12 recombinant adenovirus vector can express interleukin-12, inhibit the proliferation and induce apoptosis of ovarian cancer cells.
ABSTRACT
BACKGROUND:Gene engineering plays an important role in the process of human malignant tumor treatment, and adenovirus vectors for gene therapy are commonly used. In recent years, lactoferrin is found to exert important effects on the occurrence, development and metastasis of a variety of tumors. However, little is reported on the in vivo expression of adenovirus vector mediated lactoferrin in the human body.OBJECTIVE:To investigate the effect of adenovirus mediated human lactoferrin (Ad-hLF) on proliferation and apoptosis of cervical cancer stem-like cells.METHODS:Primary cervical cancer cells from mice were cultured in vitro to sort cervical cancer stem-like cells using SP method. Afterwards, the stem-like cells were divided into three groups, blank control group, empty vector group, and Ad-hLF group, followed by transfection with nothing, Ad-GFP and Ad-hLF, respectively. After transfection, MTT assay was used to detect the proliferation of cervical cancer stem-like cells; scratch-wound assay was employed to detect the migration of cervical cancer stem-like cells; western blot assay was used to determine the expression of lactoferrin in cervical cancer stem-like cells; and Annexin V-FITC/PI was used to detect cell apoptosis.RESULTS AND CONCLUSION:Compared with the blank control and empty vector groups, Ad-hLF significantly inhibited the proliferation of cervical cancer stem-like cells and reduced the number of scratched cells, while the cell apoptosis increased in the Ad-hLF group (P < 0.05 or P < 0.01). These findings indicate that the recombinant adenovirus vector mediated lactoferrin Ad-hLF for transfection of cervical cancer stem-like cells can be stably expressed in cervical cancer stem-like cells, and can inhibit the proliferation, increase apoptosis and reduce migration of cervical cancer stem-like cells.