+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Comparison between lentivirus and plasmid as shRNA vector targeting RhoA gene of ovary cancer cell line HO8910

, : Comparison between lentivirus and plasmid as shRNA vector targeting RhoA gene of ovary cancer cell line HO8910. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 29(5): 473-6, 480

To compare the different effects of lentivirus and plasmid as shRNA vector targeting RAS homologue gene-family member A (RhoA) of ovary cancer cell line HO8910. Lentivirus and plasmid vectors carrying siRNA targeting RhoA gene were respectively transferred into HO8910 cells. The transferred cells were screened with puromycin for the ones stably silencing RhoA gene. As passaging went on, the two types of vectors were compared in the ability of silencing RhoA gene expression by means of fluorescence microscopy, real-time quantitative PCR and Western blotting, and in the inhibitory effects on the invasion and metastasis of cells by scratch wound migration assay and tube formation assay. Two groups of stable RhoA-knockdown cell lines were established with lentivirus and plasmid as RNAi vectors, respectively. Detected by fluorescence microscopy, the expression rate of GFP decreased in the plasmid group as the culture generation increased; it was 70% and 45% at the 15th and 25th generations respectively. The expression rate of GFP in the lentivirus group maintained above 95%. Both real-time quantitative PCR and Western blotting indicated that the expression of RhoA mRNA and protein had no significant difference in the two groups at the 3rd generation; but as the culture generation increased, the expression of RhoA mRNA and protein in the lentivirus group kept at the lower level, while it increased in the plasmid group. The scratch wound migration assay and tube formation assay revealed that lentivirus as RNAi vector suppressed more stably and persistently the invasion and metastasis of ovary cancer cell line HO8910 as compared with the plasmid group (P<0.05). Lentivirus as RNAi vector can suppress the RhoA gene expression more stably as compared with the plasmid; the plasmid is more suitable for transient transfection studies.


PMID: 23643264

Other references

Maes, P.; Damart, D.; Rommens, C.; Montreuil, J.; Spik, G.; Tartar, A., 1988: The complete amino acid sequence of bovine milk angiogenin. The amino acid sequence of angiogenin isolated from bovine milk was deduced by gas-phase sequencing of the protein and its fragments. The protein contains 125 residues and has a calculated MW of 14 577. The sequence is highly homologous (65% ident...

Hueser, A.; Kloeppner, U.; Roehm, K.H.inrich, 1999: Cloning, sequence analysis, and expression of ansB from Pseudomonas fluorescens, encoding periplasmic glutaminase/asparaginase. A gene (ansB) encoding a class II glutaminase/asparaginase has been cloned from Pseudomonas fluorescens and characterized by DNA sequencing, promoter analysis and heterologous expression in Escherichia coli. We show that ansB is monocistronic and...

Braekevelt C.R., 1975: The fine structure of the retinal pigment epithelium in caiman sclerops. Anatomical Record 181(2): 317

Barnes, H., 1916: On Anthony Askew, M.D., F.R.S., and his Library. Proceedings of the Royal Society of Medicine 9(Sect Hist Med): 23-27

Vugia, D.J.; Holmberg, E.; Steffe, E.M.; Ascher, M.S.; Gallo, D., 1996: A human case of monocytic ehrlichiosis with adult respiratory distress syndrome in northern California. Western Journal of Medicine 164(6): 525-528

Khole, V.; Hegde, U., 1992: Monoclonal anti-idiotypic antibody to progesterone with internal image properties. An auto-anti-idiotypic approach was used to generate mouse monoclonal anti-idiotypic antibody E9F11F6 (Ab2) to progesterone. A conjugate of 4-pregnane 3,20 dione and bovine serum albumin was used as the immunogen. E9F11F6 bound to a rabbit anti-pr...

Anonymous, 1944: Choice of species. Species of Eucalyptus, particularly E. rostrata and E. robusta, are considered to be far more satisfactory as plantation trees than Callitris glauca, which in the past has been a favourite species for this purpose in the Northern Provinces but has...

Lesina, V.; Garkina, G., 1976: Pests of vegetable crops in greenhouses during the fruiting period. Kartofel' i ovoshchi: 6 31-32

Katzke D.; Steinbach E., 1985: Experimental sialadenosis. Archives of Oto-Rhino-Laryngology 241(2): 121

Drucker D.J.; Philippe J.; Jepeal L.; Habener J.F., 1987: Enhancer like sequences mediate islet cell specific glucagon gene transcription. Clinical Research 35(3): 657A