Construction of expression vector for RNA interference of hypoxia-inducible factor 1alpha and its function in the study of prostatic carcinoma cell line PC-3

• Published in: Sheng wu yi xue gong cheng xue za zhi Vol. 28; no. 1; p. 129
• Main Authors: Lin, Rongfu, Geng, Aimin, Li, Bo'an
• Format: Journal Article
• Language: Chinese
• Published: China 01.02.2011
• Subjects: Base Sequence; Genetic Vectors; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Hypoxia-Inducible Factor 1, alpha Subunit - genetics; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Male; Molecular Sequence Data; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Prostatic Neoplasms - pathology; RNA Interference; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transfection
• ISSN: 1001-5515

According to the specific construction of the pSUPER. retro. puro vector, RNA interfering with the hypoxia-inducible factor 1alpha (HIF-1alpha) plasmid was constructed in this research programme. The efficiency was (87.15 +/- 2.36)% and green fluorescent protein was observed after 36 hours when the constructed plasmid co-transformed into the prostatic carcinoma cell line PC-3; compared with the control groups, this constructed plasmid could reduce the expression of total RNA and protein in PC-3 cells significantly after 48 hours. The cells were checked out by the plasmid resistance against the puromycin biotin, the clones of which were selected and enlarged for two weeks, then the cell strain of pSUPER-siHIF-1alpha/PC-3 was collected. The HIF-1alpha protein expression of the pSUPER-siHIF-1alpha/ PC-3 cells of also decreased significantly. The results showed that the RNA interference could be used in the construction of expression vector of constructed siRNA inhibiting the expression of HIF-1alpha with PC-3 ce