Antitumor activity of anti-type IV collagenase monoclonal antibody and its lidamycin conjugate against colon carcinoma

AIM： Type IV collagenase including MMP-2 and -9 plays an important role in cancer cell invasion and metastasis and is an attractive target for rnAb-directed therapy. The irnrnunoreactivity of rnAb 3G11, a rnAb directed against type Ⅳ collagenase in human colorectal carcinomas, was studied by irnrnuno-histochernical （IHC） staining, rnAb 3G11 was conjugated to an antiturnor antibiotic lidarnycin （LDM）. The antiturnor activity of 3G11-LDM conjugate against colon carcinoma was investigated in mice. METHODS： ELISA, gelatin zyrnography, and Western blot assay were used for the biological characterization of rnAb 3G11. The irnrnunoreactivity of rnAb 3G11 with human colorectal carcinomas was detected by IHC staining. The cytotoxicity of LDM and 3G11-LDM conjugate to human colon carcinoma HT-29 cells was examined by clonogenic assay and MTT assay. The therapeutic effect of conjugate 3G11-LDM was evaluated with colon carcinoma 26 in mice. RESULTS： As shown in ELISA, mAb 3Gll reacted specifically with type IV collagenase, while 3G11-LDM conjugate also recognized specifically its respective antigen. In IHC assay, mAb 3G11 showed positive irnrnunoreactivity in most cases of colorectal carcinoma, and negative irnrnunoreactivity in the adjacent non-malignant tissues. By gelatin zyrnography, the inhibition effect of rnAb 3G11 on the secretion activity of type IV collagenase was proved. In terms of IC50 values in MTT assay, the cytotoxicity of LDM to human colon carcinoma HT-29 cells was 10 000-fold more potent than that of rnitornycin C （MMC） and adriarnycin （ADM）. 3G11- LDM conjugate also displayed extremely potent cytotoxicity to human colon carcinoma HT-29 cells with an IC50 value of 5.6× 10^-19 mol/L. 3G11-LDM conjugate at the doses of 0.05 and 0.1 mg/kg inhibited the growth of colon carcinoma 26 in mice by 70.3 and 81.2%, respectively. CONCLUSION： mAb 3G11 is immunoreactive with human colorectal carcinoma and its conjugate with LDM is highly effective against colon carcinoma in mice.

瞄准：包括 MMP-2 和 -9 的类型 IV 胶原酶在癌症房间侵略和转移起一个重要作用并且是为指导 mAb 的治疗的一个吸引人的目标。mAb 3G11 的免疫反应，在人对类型 IV 胶原酶指导的 mAb 渲染表面的癌，被学习由免疫组织化学(IHC ) 染色。mAb 3G11 被结合到反肿瘤抗菌素 lidamycin (LDM ) 。对冒号癌的 3G11-LDM conjugate 的反肿瘤活动在老鼠被调查。方法：ELISA，明胶 zymography，和西方的污点试金被用于 mAb 3G11 的生物描述。有人的 mAb 3G11 的免疫反应渲染表面的癌被染色的 IHC 检测。到人的结肠癌 HT-29 房间的 LDM 和 3G11-LDM conjugate 的细胞毒性被 clonogenic 试金和 MTT 试金检验。conjugate 3G11-LDM 的治疗学的效果与在老鼠的冒号癌 26 被评估。结果：是出现在 ELISA， mAb 3G11 与类型 IV 胶原酶明确地反应了，当 3G11-LDM conjugate 也明确地认出了它的各自的抗原时。在 IHC 试金， mAb 3G11 在颜色的大多数情况中显示出积极免疫反应表面的癌，和在邻近的非恶意的纸巾的否定免疫反应。由明胶 zymography，在类型 IV 胶原酶的分泌物活动的 mAb 3G11 的抑制效果被证明。以在 MTT 试金的 IC50 价值，到人的结肠癌 HT-29 房间的 LDM 的细胞毒性更是 10,000 褶层比丝裂霉素Ｃ(MMC ) 和 adriamycin (ADM ) 的有势力。3G11-LDM conjugate 也极其与 5.6 x 10 的 IC50 价值显示了有势力细胞毒性到人的冒号癌 HT-29 房间(-19) mol/L。在 0.05 和 0.1 mg/kg 的剂量的 3G11-LDM conjugate 分别地在 70.3 和 81.2% 禁止了在老鼠的结肠癌 26 的生长。结论：mAb 3G11 是与有 LDM 的表面的癌和它的 conjugate 是的人的颜色反应的免疫对在老鼠的冒号癌高度有效。