Journal: Molecular medicine reports

Article Title: Antitumor activity of an adenovirus harboring two therapeutic genes, anti-VEGF ribozyme and human IL-24, in colon cancer.

doi: 10.3892/mmr_00000158

Figure Lengend Snippet: Figure 1. Schematic representation of the recombinant adenoviral vectors. An expression cassette containing (A) anti-VEGF ribozyme (Rz), (B) IL-24 and (C) anti-VEGF ribozyme + IL-24 (Rz/IL-24) was inserted into the deleted E1 region.

Article Snippet: The amplification conditions consisted of an initial 5-min denaturation step at 94 ̊C, followed by 50 cycles of 95 ̊C for 15 sec and 60 ̊C for 60 sec. To quantify the secreted VEGF protein in conditioned media from cells, human VeGF eliSa was performed using a human VeGF eliSa Kit (Boster, china) following the manufacturer's protocol with 100-μl samples diluted 1:50.

Techniques: Recombinant, Expressing

Journal: Molecular medicine reports

Article Title: Antitumor activity of an adenovirus harboring two therapeutic genes, anti-VEGF ribozyme and human IL-24, in colon cancer.

doi: 10.3892/mmr_00000158

Figure Lengend Snippet: Figure 2. Expression of anti-VEGF ribozyme RNA and IL-24 mRNA and protein in viral infected HT-29. A: (a) RT-PCR demonstrated that HT-29 cells did not express IL-24. Lane 1, marker; lane 2, cDNA from untreated HT-29 cells where no IL-24 expression was noted; lane 3, negative control, PCR for plasmid of pTrack-CMV-Rz; and lane 4, positive control, PCR for plasmid of pTrack-CMV-IL-24. A 621-bp DNA fragment was noted. (b) RT-PCR analysis of the expres sion of anti-VEGF ribozyme and IL-24 RNA in HT-29 cells. Lane 1, IL-24 in cells infected with Ad-IL-24; lane 2, anti-VEGF ribozyme with an expected length of 162 bp in cells infected with Ad-Rz; lane 3, marker; lane 4, IL-24 in cells infected with Ad-Rz/IL-24; and lane 5, anti-VEGF ribozyme in cells infected with Ad-Rz/IL-24 with an expected length of 214 bp. B: Results of immunofluorescence staining of IL-24 protein expression in cells treated with (c) PBS, (d) Ad-IL-24 and (e) Ad-Rz/IL-24. Left, light microscopy; right, fluorescence microscopy.

Article Snippet: The amplification conditions consisted of an initial 5-min denaturation step at 94 ̊C, followed by 50 cycles of 95 ̊C for 15 sec and 60 ̊C for 60 sec. To quantify the secreted VEGF protein in conditioned media from cells, human VeGF eliSa was performed using a human VeGF eliSa Kit (Boster, china) following the manufacturer's protocol with 100-μl samples diluted 1:50.

Techniques: Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Marker, Negative Control, Plasmid Preparation, Positive Control, Immunofluorescence, Staining, Light Microscopy, Fluorescence, Microscopy

Journal: Molecular medicine reports

Article Title: Antitumor activity of an adenovirus harboring two therapeutic genes, anti-VEGF ribozyme and human IL-24, in colon cancer.

doi: 10.3892/mmr_00000158

Figure Lengend Snippet: Figure 3. Cells treated with Ad-Rz/IL-24, Ad-IL-24 and Ad-Rz expressed reduced levels of VEGF. (A) Real-time PCR results, (B) ELISA results of VEGF protein secretion.

Article Snippet: The amplification conditions consisted of an initial 5-min denaturation step at 94 ̊C, followed by 50 cycles of 95 ̊C for 15 sec and 60 ̊C for 60 sec. To quantify the secreted VEGF protein in conditioned media from cells, human VeGF eliSa was performed using a human VeGF eliSa Kit (Boster, china) following the manufacturer's protocol with 100-μl samples diluted 1:50.

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay