Nishimura Mane, Shimizu Yuki, Fujii Tomohiro, Okada Yu, Yamamoto Takeshi, Ogawa Yoshimasa, Koi Toru, Suehiro Yutaka, Yamasaki Takahiro, Nishikawa Jun
Faculty of Laboratory Science, Yamaguchi University Graduate School of Medicine, Ube, Japan.
Ushio Inc., Chiyoda-ku, Tokyo, Japan.
Sci Rep. 2025 May 14;15(1):16787. doi: 10.1038/s41598-025-00477-6.
Ultraviolet (UV) light is never used for gene transfer because it damages DNA and harms cellular and plasmid DNA. A light source selectively radiating Far UV-C (F-UV) light causes less DNA damage. We investigated potential introduction of plasmids into cells by irradiating them with F-UV light. COS-7 and CHO-K1 cells were irradiated with 222 nm F-UV light. Then, DNA solution containing green fluorescent protein (EGFP) plasmid was added to the culture and incubated (37 °C, 5% CO, 24 h) for fluorescence microscopy. Cytotoxicity of cells irradiated with the same energy as for gene transfer was evaluated. Characteristics of EGFP-positive cells were compared with non-transfected cells by propidium iodide (PI) and Hoechst staining. Cells with distinct green fluorescence were observed after F-UV light irradiation and addition of 200 ng of EGFP plasmid. COS-7 cells showed the highest number of EGFP-positive cells at 0.5 mJ/cm irradiation, whereas that for CHO-K1 cells was at 1 mJ/cm irradiation. Cytotoxicity was low in COS-7 at ≤ 1 mJ/cm irradiation and CHO-K1 at ≤ 0.5 mJ/cm. Transfected cells did not incorporate PI, and their nuclei did not differ from those of non-transfected cells. We successfully transfected two cell lines with EGFP plasmids by F-UV irradiation.
紫外线（UV）从不用于基因转移，因为它会损伤DNA并损害细胞和质粒DNA。选择性辐射远紫外线C（F-UV）的光源造成的DNA损伤较小。我们研究了用F-UV光照射将质粒导入细胞的可能性。用222nm的F-UV光照射COS-7和CHO-K1细胞。然后，将含有绿色荧光蛋白（EGFP）质粒的DNA溶液加入培养物中，并在（37°C，5%CO₂，24小时）条件下孵育，用于荧光显微镜观察。评估了与基因转移相同能量照射的细胞的细胞毒性。通过碘化丙啶（PI）和Hoechst染色，将EGFP阳性细胞的特征与未转染细胞进行比较。在用F-UV光照射并添加200ng EGFP质粒后，观察到具有明显绿色荧光的细胞。在0.5mJ/cm²照射下，COS-7细胞显示出最高数量的EGFP阳性细胞，而CHO-K1细胞在1mJ/cm²照射下数量最多。在≤1mJ/cm²照射下，COS-7细胞的细胞毒性较低，在≤0.5mJ/cm²照射下，CHO-K1细胞的细胞毒性较低。转染细胞不摄取PI，其细胞核与未转染细胞的细胞核没有差异。我们通过F-UV照射成功地用EGFP质粒转染了两种细胞系。
