抗Ⅳ型胶原酶单抗3G11及其F(ab′)_2与Fab′片段的抗肿瘤作用

Antitumor Effects of Anti-type IV Collagenase Monoclonal Antibody 3G11 and Its F(ab′)_2 and Fab′ fragments.

目的研究抗Ⅳ型胶原酶单抗3G11及其F(ab′)2、Fab′片段对小鼠肝癌22(H22)及小鼠Lewis癌肺转移的治疗作用。方法ELISA实验检测3G11及其片段与多种肿瘤细胞及Ⅳ型胶原酶的免疫反应性,明胶酶谱法测定其对HT-1080细胞分泌Ⅳ型胶原酶活性的影响,MTT法测定其对H22细胞的细胞毒作用。用小鼠肝癌22及小鼠Lewis肺癌模型评价单抗3G11及其片段抑制肿瘤生长及抗肿瘤转移活性。结果单抗3G11及其片段对多种肿瘤细胞及Ⅳ型胶原酶均呈阳性反应,单抗3G11抑制肿瘤细胞HT-1080分泌Ⅳ型胶原酶的活性。单抗3G11对H22细胞有微弱的细胞毒作用。单抗3G1130mg/kg对小鼠移植性肝癌的抑制率为51·8%,F(ab′)210mg/kg为49·9%,Fab′20mg/kg为39·3%。F(ab′)2片段10mg/kg对小鼠Lewis癌肺转移的抑制率为76%。结论抗Ⅳ型胶原酶单抗3G11及其片段对小鼠移植性肝癌22生长以及对小鼠Lewis癌肺转移有中度抑制作用,可能用于研制抗体靶向药物。

Objective To study the antitumor effects of the monoclonal antibody 3G11 and its fragment in vivo against hepatoma 22 tumor model and Lewis lung carcinoma model in mice.Methods Immunoreactivity ofmAb 3G11 and its F （ab＇）2 and Fab＇ fragments to type Ⅳ collagenase and a series of tumor cells was determined by ELISA. Gelatin - zymography analyses were used to detect the binding characteristics of mAb 3G11, and the cytotoxicity was examined by clonogenic assay. Antitumor effects of mAb 3G11 and its fragment in vivo were evaluated using mouse hepatoma 22 tumor model and Lewis lung carcinom model in mice.Results mAb 3G11 and its fragment showed high specificity for their target antigens by ELISA. mAb 3G11 and its F （ab＇）2 fragment markedly inhibited the gelatinase activity in a dose dependent manner and no obvious cytotoxicity to KB and BEL- 7402 cells was observed.At dosage of 30 mg/kg, mAb 3G11 inhibited the growth of hepatoma 22 tumor by 51.8%, and F （ab＇）2 fragment 10 mg/kg by 49.9%, and Fab＇ fragment 20 mg/kg by 39.3%, respectively. At dose of 10 mg/kg, F （ab＇）2 suppressed the metastases of Lewis lung carcinoma by 76 %. Conclusion Anti - type Ⅳ collagenase mAb 3G11 and its fragments moderately inhibits tumor growth of hepatoma 22 and metastasis of Lewis lung carcinoma in mice. It might be useful for manufacture of antibody - based therapeutics.