CAJ | 학술논문

利用大肠杆菌表达系统制备了重组融合蛋白antiEGFR/MEL,并用Ni2+层析柱对其进行了纯化。该重组蛋白中抗表皮生长因子受体(antiEGFR)单链抗体(scFv)主要靶向喉癌细胞中的EGFR,而蜂毒肽(MEL)主要抑制肿瘤细胞增殖。采用SDS-PAGE和Western blot检测证明了antiEGFR/MEL的有效表达。共聚焦显微镜和流式细胞术实验结果表明, antiEGFR/MEL可与Hep-2肿瘤细胞有效结合,而几乎不与EGFR阴性的Jurkat细胞结合。噻唑蓝( MTT)检测结果说明, antiEGFR/MEL可有效抑制人喉癌细胞Hep-2的增殖。以上结果表明, antiEGFR/MEL能够有效靶向EGFR阳性肿瘤细胞,并有效抑制肿瘤细胞增殖,有望应用于EGFR靶向肿瘤治疗。
이용대장간균표체계통제비료중조융합단백antiEGFR/MEL,병용Ni2+층석주대기진행료순화。해중조단백중항표피생장인자수체(antiEGFR)단련항체(scFv)주요파향후암세포중적EGFR,이봉독태(MEL)주요억제종류세포증식。채용SDS-PAGE화Western blot검측증명료antiEGFR/MEL적유효표체。공취초현미경화류식세포술실험결과표명, antiEGFR/MEL가여Hep-2종류세포유효결합,이궤호불여EGFR음성적Jurkat세포결합。새서람( MTT)검측결과설명, antiEGFR/MEL가유효억제인후암세포Hep-2적증식。이상결과표명, antiEGFR/MEL능구유효파향EGFR양성종류세포,병유효억제종류세포증식,유망응용우EGFR파향종류치료。
Laryngeal carcinoma is poor prognosis and patients with laryngeal carcinoma usually present late leading to the reduced treatment efficacy and high rate of recurrence. Incidence and mortality rates of laryngeal carcinoma are equivalent, suggesting a failure of current therapies, despite the variety of combined modality approaches which have been introduced to complement surgical treatment. A successful cancer therapeutic should target tumours specifically with limited systemic toxicity. Herein, the recombinant antiEGFR/MEL pro-tein was expressed in Escherichia coli( E. coli) , refolded and purified on an immobilized Ni2+-affinity chroma-tography column. In the protein, anti epidermal growth factor receptor( EGFR) single chain antibody was used to target the EGFR in the laryngeal cancer cell, and the melittin was used to mediate inhibition of cell growth. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis( SDS-PAGE) and Western blotting analysis revealed that antiEGFR/MEL was sufficiently expressed. Confocal microscopy and flow cytometry demonstrated that antiEGFR/MEL bound specifically to Hep-2 cells, as almost no binding to Jurkat cells was observed under identical time and dosage conditions. MTT assay showed that antiEGFR/MEL suppressed the growth of Hep-2 cells effecitively. Collectively, these results suggest that antiEGFR/MEL is biologically active and specific toward EGFR-positive tumor cells and may represent an effective EGFR-targeted cancer therapy.