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Photodynamic antisense regulation of human cervical carcinoma cell growth using psoralen-conjugated oligo(nucleoside
A Murakami1, A Yamayoshi, R Iwase
1Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, 606-8585, Kyoto, Japan. akiram@ipc.kit.ac.jp
Abstract:
The antisense strategy has been applied to regulate gene expression in a sequence specific manner, which enables suppression of the proliferation of cancer cells and exploration of the functions of unknown genes. In order to generalize and to enhance the ability of the strategy, functionalization of antisense DNAs was done using a photo-crosslinking reagent, 4,5',8-trimethylpsoralen, and the possibility of photodynamic antisense regulation of gene expression was examined. Psoralen-conjugated oligo(nucleoside phosphorothioate)s (Ps-S-oligo) were prepared and used to inhibit the proliferation of human cervical carcinoma cells. Upon UVA irradiation of Ps-S-oligo treated cells, Ps-S-oligo complementary to the initiation codon region (Ps-P-As) of HPV18-E6*-mRNA of human cervical carcinoma cells inhibited drastically the cell growth (IC(50)=16 nM). In contrast, Ps-S-oligo with mismatched sequences and scrambled one showed lesser inhibitory effects than Ps-P-As. These results showed that the inhibition by Ps-S-oligo was dependent on (a) sequence, (b) UVA irradiation, (c) concentration and (d) cell line. The amount of intact HPV18-E6*-mRNA was decreased in a sequence dependent manner, indicating that the antiproliferative effect of Ps-P-As was an antisense manner. The psoralen-conjugated antisense DNA has significant potential to regulate gene expression, which may provide useful information to explore the novel gene regulating reagents.
Insights
Functionalized antisense DNA (Ps-S-oligo) effectively inhibits human cervical carcinoma cell growth via photodynamic regulation. This sequence-specific approach, activated by UVA light, offers a novel strategy for gene expression control and cancer therapy development.
Area of Science:
- Molecular Biology
- Antisense Technology
- Photodynamic Therapy
Background:
- Antisense strategies regulate gene expression for cancer research and gene function studies.
- Enhancing antisense DNA efficacy and generalizability is crucial for broader applications.
Purpose of the Study:
- To investigate the photodynamic antisense regulation of gene expression using psoralen-conjugated antisense DNA (Ps-S-oligo).
- To evaluate the antiproliferative effects of Ps-S-oligo on human cervical carcinoma cells.
Main Methods:
- Preparation of psoralen-conjugated oligo(nucleoside phosphorothioate)s (Ps-S-oligo).
- Treatment of human cervical carcinoma cells with Ps-S-oligo and subsequent UVA irradiation.
- Assessment of cell growth inhibition and HPV18-E6*-mRNA levels.
Main Results:
- Ps-S-oligo complementary to HPV18-E6*-mRNA (Ps-P-As) significantly inhibited cancer cell proliferation (IC(50)=16 nM) upon UVA irradiation.
- Inhibition was sequence-dependent, requiring UVA irradiation, specific concentration, and cell line.
- HPV18-E6*-mRNA levels decreased in a sequence-dependent manner, confirming antisense activity.
Conclusions:
- Psoralen-conjugated antisense DNA demonstrates significant potential for sequence-specific gene expression regulation.
- This photodynamic approach offers a novel avenue for developing gene-regulating reagents and potential cancer therapies.