モノクロナ-ル抗体,遺伝子を用いた耐性癌の基礎的な診断と治療に関する研究

利用单克隆抗体和基因进行耐药癌症基础诊断和治疗的研究

• 批准号: 03151028
• 负责人: 白川 茂
• 金额: $ 9.34万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Cancer Research
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03151028/
• 关键词: 多剤耐性腫瘍; P糖蛋白(Pーglycoprotein;Pーgp); 抗Pーgp抗体MRK16; 抗Pーgp抗体C219; 多剤耐性遺伝子(mdr1); mdr1mRNA発現; ヒトマウスキメラ抗体; magneticーaffinity cell sorring(MACS)法

(1)基礎的に抗P糖蛋白(Pーgp)抗体MRK16(鶴尾)の反応性とmdr1遺伝子のmRNA発現程度を培養細胞株で検討し,中等度以上のmRNA発現細胞株でMRK16陽性であった(下山)。(2)ヒト腫瘍nude mouse移植株47株のうち多剤耐性を示す33株中21株でRTーPCR法によりmdr1遺伝子の発現を認め,このうち8株が抗Pーgp抗体C219陽性であった。なおMRK16とC219による固型腫瘍材料の免疫染色至適条件が決定された(玉置)。(3)臨床材料につきmdr1遺伝子発現を6種類のamplimerを用いてRTーPCR法で検索し,amplimerは200〜300bp前後のものが感度が高く,少量の試料で解析可能であった(向山)。(4)白血病,悪性リンパ腫などの造血器腫瘍でのPーgpの関与はin vivoの耐性細胞株で確認された。in vivoの症例検索がなされ,128例の造血器腫瘍でMRK16 F(ab')_2抗体(鶴尾)を用いた間接法で11例陽性,直接法では3例(リンパ腫再発例)の化学療法不応例のみに陽性であった(上田)。また64例の各種白血病症例で同上抗体でのFACS解析陽性例は8例で,うち5例は単球性性格を示し,経過を追跡し得た治療抵抗性8例でmdr1遺伝子発現を経過中に認めたものはPh(+)ALL,MDS各1例で臨床上検出限界に問題が残った(白川)。(5)固型癌については,MRK16の肺癌培養細胞42株の検討では全例陰性であった(上田)が,C219の免疫染色で肺癌,転移性肺腫瘍76例中26例(24%)に陽性(広橋),大腸癌ではC219で高率(68%)に陽性(玉置),また大腸癌,未分化甲状腺癌,肝癌,胎盤など(菅原),乳癌,卵巣癌など(向山)でPTーPCR法によるmdr1mRNAの有意な発現があり,化学療法抵抗性と関連があった。(6)MRK16とMRK17はADCC誘導に際し,リンパ球はMRK16においてのみeffectorとして,単球は両抗体において働くことが判明した。またヒトマウスキメラ抗体の臨床応用への可能性が示唆された(曽根,鶴尾)。(7)MRK16とmagnetic particleをcouplingしたbeadsを用いるmagneticーaffinity cell sorting(MACS)法が開発され,Pーgp発現多剤耐性細胞の分離に成功した。

(1)The basic anti-P glycoprotein (P gp) antibody MRK16 (Tsuru) was found to be resistant to mdr1 mRNA expression in cultured cell lines, and MRK16 positive in cell lines with moderate or higher mRNA expression. (2)Among the 47 transplanted strains, 21 strains were found to be resistant to multiple enzymes by RT PCR, and 8 strains were positive for anti-P-gp antibody C219. MRK16 and C219 were used to determine the appropriate immunostaining conditions for solid tumor materials. (3)6 kinds of amplimer were detected by RT PCR in clinical materials. The sensitivity of amplimer around 200 ~ 300bp was high, and a small number of samples could be analyzed. (4)The relationship between P gp and in vivo resistant cell lines in leukemia, leukemia, and hematopoietic tumors was confirmed. In vivo, MRK16F (ab')_2 antibody was detected in 128 cases of hematopoietic tumors. 11 cases were positive by indirect method, 3 cases were positive by direct method, and no cases were positive by chemotherapy. FACS analysis of 64 leukemia cases showed positive antibodies in 8 cases, negative in 5 cases, negative in 8 cases, negative in 5 cases, negative in 8 cases, negative in 1 case, negative in 1 case. (5)42 strains of MRK16 lung cancer cells were negative in all cases,C219 immunostaining was positive in 26 cases (24%) of 76 cases of lung cancer, metastatic lung cancer, colorectal cancer, undifferentiated thyroid cancer, liver cancer, placental carcinoma, breast cancer, breast cancer, breast cancer, The expression of mdr1 mRNA in ovarian cancer by PCR is related to chemotherapy resistance. (6)MRK 16 and MRK17 were identified as ADCC inducing factors. The possibility of clinical use of antibodies is demonstrated. (7)MRK16 and magnetic particle coupling beads were successfully isolated by magnetic affinity cell sorting(MACS) method.

项目成果

Maeda K,et al: "A novel differentiation antigen on human monocytes that is specifically induced by granulocyte-macrophage colony-stimulating factor ar 1L-3" J.Immunol. 146. 3779-3784 (1991)

Maeda K 等人：“一种新型人单核细胞分化抗原，由粒细胞-巨噬细胞集落刺激因子 ar 1L-3 特异性诱导”J.Immunol。

Sugawara I,et al: "Characterization of an etoposide-resistant human K562 cell line,K/eto." Jpn.J.Cancer Res.82. 1035-1043 (1991)

Sukawara I 等人：“依托泊苷抗性人 K562 细胞系 K/eto 的表征。”

Nishioka Y,et al: "Effector cell analysis of human multidrug-resistant cell killing by mouse-human chimeric antibody against P-glycoprotein" Jpn.J.Cancer Res.

Nishioka Y 等人：“针对 P-糖蛋白的小鼠-人嵌合抗体杀伤人类多重耐药细胞的效应细胞分析”Jpn.J.Cancer Res。

Sekido Y,et al: "Preferential expression of c-kit protooncogene transcripts in small cell lung cancer." Cancer Research. 51. 2416-2419 (1991)

Sekido Y 等人：“c-kit 原癌基因转录物在小细胞肺癌中的优先表达。”

Shimoyama Y,et al: "Expression of E-and P-cadherin in gastric carcinomas." Cancer Research. 51. 2185-2192 (1991)

Shimoyama Y 等人：“E-和 P-钙粘蛋白在胃癌中的表达”。