Cat.No. : LVIM022Z
Titer: ≥1*10^7 TU/mL / ≥1*10^8 TU/mL / ≥1*10^9 TU/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | LVIM022Z |
| Description | Lentivirus expressing untagged HPV16 E6-E7 under the control of CMV promoter. This lentivirus contains no antibiotic selection marker. |
| Target Gene | E6-E7 |
| Species | HPV16 |
| Titer | Varies lot by lot, for example, ≥1*10^7 TU/mL, ≥1*10^8 TU/mL, ≥1*10^9 TU/mL etc. |
| Size | Varies lot by lot, for example, 100 ul, 500 ul, 1 mL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality lentivirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between lentivirus particle lots. | |
| Mycoplasma | Creative Biogene routinely tests for mycoplasma contamination using a mycoplasma detection kit. Cell lines are maintained for approximately 20 passages before being discarded and replaced with a new vial of early passage cells. Approximately 2 weeks after thawing, cell culture supernatants are tested for mycoplasma contamination. Creative Biogene ensures that lentiviral products are free of mycoplasma contamination. |
| Purity | Creative Biogene evaluates the level of impurities, such as residual host cell DNA or proteins, in prepared lentiviral vectors to ensure they meet quality standards. |
| Sterility | The lentiviral samples were inoculated into cell culture medium for about 5 days and the growth of bacteria and fungi was tested. Creative Biogene ensures that the lentiviral products are free of microbial contamination. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of lentivirus to deliver genetic material into target cells, and assess gene expression and functional activities. |
| Proviral Identity Confirmation | All Creative Biogene lentiviral vectors are confirmed to have correctly integrated provirus using PCR. This test involves transducing cells with serial dilutions of the lentiviral vector, harvesting the cells a few days later, and isolating genomic DNA. This DNA is then used as a template to amplify a portion of the expected lentiviral insert. |
High-risk human papillomavirus (HPV), especially HPV16, is closely associated with the occurrence of human esophageal squamous cell carcinoma (ESCC). Here, the researchers aimed to investigate the potential role and mechanism of HPV16 in the occurrence and development of ESCC. Subsequent studies have shown that ESCC cells stably transfected with HPV16 E6-E7 lentiviral vectors exhibited obvious cancer stem cell-like cell (CSCs) phenotypes, such as migration, invasion, sphere formation, high expression of p75NTR, a CSCs marker in ESCC, chemotherapy resistance, radioresistance, in vitro anti-apoptosis ability and in vivo carcinogenicity. HPV16 E6-E7 induced activation of the PI3K/Akt signaling pathway, and this effect could be effectively inhibited by the specific PI3K inhibitor LY294002. It was also shown that inhibition of the PI3K/Akt signaling pathway by PI3K and Akt siRNA could reverse the effects induced by HPV16 E6-E7 in ESCC cells. In conclusion, this study demonstrates that HPV16 E6-E7 promotes the CSCs phenotype in ESCC cells by activating the PI3K/Akt signaling pathway. Targeting the PI3K/Akt signaling pathway in HPV16-positive tissues is an effective therapeutic approach for ESCC patients.
HPV16 E6-E7 protein was stably expressed in ESCC cells transfected with HPV16 E6-E7 lentiviral vectors, which were labeled as Eca109-psb and TE-1-psb cells, respectively; no expression of HPV16 E6-E7 protein was observed in ESCC cells transfected with control vectors, which were labeled as Eca109-control and TE-1-control cells, respectively. The transfection efficiency was detected and confirmed by Western blotting (Figure 1A). After 72 h of experimental treatment of Eca109 and TE-1 cells, the transfection results of HPV16 E6-E7 lentiviral vectors were observed by fluorescence microscopy (Figure 1B). The migration and invasion abilities of ESCC cells were detected by Transwell assay, and the results are shown in Figures 1C and 1E, and quantitative analysis was performed (Figures 1D and 1F). Compared with control cells, the migration and invasion abilities of Eca109-psb cells were significantly improved in the 24h (without matrigel) migration assay and the 48h (with matrigel) invasion assay, indicating that HPV16 E6-E7 can enhance the migration and invasion abilities of ESCC cells in vitro.
Figure 1. HPV16 E6-E7 lentiviral vector stably transfected ESCC cells. (Xi R, et al., 2016)
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