Basic study on chemotherapy combined with gene transfection for head and neck cancer

化疗联合基因转染治疗头颈癌的基础研究

• 批准号: 10470354
• 负责人: SAITO Hitoshi
• 金额: $ 4.42万
• 依托单位: Fukui Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470354/
• 关键词: Head and neck cancer; Apoptosis; bax; p53; bcl-2; CAD; Fas-Fas ligand; Chemotherapy; 頭頚部癌; 遺伝子治療; Fas; Fasリガンド; CDDP; 遺伝子銃; CDDP耐性; グルタチオン; MRP; Bax; Bcl-2; 頭頸部癌; 抗癌剤感受性試験; 血小板由来増殖因子; Tunicamycin; 顆粒球刺激因子; Bax遺伝子

The chemosensitive rate of 139 cases of head and neck cancer evaluated by ATP method was in the order of 5-FU (27%) >CDDP=CBDCA>MTX>PEP (23%). Therefore, the purpose of this study is to conquer the chemoresistance in combination with gene transfection relating to apoptosis.1998 : Among the above 5 sensitive drugs, CDDP was employed for the further study, because CDDP is one of the apoptosis-promoting anticancer drugs. Apoptosis by CDDP is regulated with glutathione and with tunicamycin, an inhibitor of glycosylation. A correlation between Bax expression and 5-year survival rate in 57 cases of oropharyngeal squamous cell carcinomas was investigated by immunohistochemical analysis. The cases expressed high apoptosis-promoting gene bax showed significantly better 5-year survival rate.1999 : Overexpression of p53 showed significantly poor prognosis, while that of Bcl-2 did not show any correlation to it. Correlation between the expressions of p53, Bax, Bcl-2 and apoptotic ratio evaluated b … More y TUNEL staining did not show significant relation, but Bax-positive group showed higher tendency to apoptosis. Since clinical cases highly expressed gene bax showed better prognosis, it was investigated whether the head and neck cancer cell line and CDDP-resistant cell line transfected gene bax by electroporation become more sensitive to CDDP in vitro. The cell lines transfected bax showed significantly higher sensitivity to CDDP.A cancer cell line expressing bcl-2 showed significantly increased CDDP sensitivity by its antisense.2000 : In vivo study on chemotherapy with some genes promoting apoptosis was carried out.(1) Anticancer effect of apoptosis-promoting bax gene and its combined effect with CDDP were investigated. Percutaneous transfer of bax gene by particle-mediated delivery (gene gun system) inhibited the growth of mouse CDDP-resistant SCC.Furthermore, a combination with bax gene and CDDP at 3-day intervals markedly inhibited the growth of mouse SCC (p<0.0001).(2) Anticancer effect of CAD (caspase-activated DNase) and its combined effect with CDDP were investigated. The final DNA fragmentation during apoptosis is controlled by CAD.Although percutaneous transfer of CAD gene alone delivered by the gene gun did not show clear inhibition, a combination with CAD and CDDP significantly inhibited the growth of SCC (p<0.05).(3) Anticanacer effect of Fas-Fas ligand system was investigated. Since the C3H mouse SCC did not express Fas ligand, this ligand was delivered into the SCC by the same gene gun, and examined the anticancer effect. The growth of SCC transfected Fas ligand was significantly inhibited (p<0.0003), and also significant elongation of the survival time was observed.Among the above 3 apoptosis-promoting gene transfections, the anticancer effect of bax gene +CDDP seemed to be most effective. Less

139例头颈部恶性肿瘤的ATP敏感率依次为5-FU（27%）>CDDP=CBDCA>MTX>PEP（23%）。因此，本研究的目的是克服化疗耐药性与基因转染相关的乳腺癌。1998年：在上述5种敏感药物中，CDDP被用于进一步研究，因为CDDP是促进乳腺癌的抗癌药物之一。CDDP引起的细胞凋亡受谷胱甘肽和衣霉素（一种糖基化抑制剂）的调节。应用免疫组化方法检测57例口咽鳞癌组织中Bax蛋白表达与5年生存率的关系。促凋亡基因bax高表达者5年生存率明显提高。1999年：p53过表达者预后明显差，Bcl-2表达与预后无相关性。B评价p53、Bax、Bcl-2表达与凋亡率的相关性 ...更多信息 γ TUNEL染色显示无明显相关性，但TGF β 1阳性组有更高的凋亡倾向。由于临床病例高表达基因bax显示更好的预后，因此研究了通过电穿孔转染基因bax的头颈癌细胞系和CDDP抗性细胞系是否在体外对CDDP变得更敏感。bax基因转染的肿瘤细胞对顺铂的敏感性显著提高，bcl-2基因反义表达的肿瘤细胞对顺铂的敏感性显著提高。2000年：利用促凋亡基因进行化疗的体内研究。(1)探讨促凋亡基因bax的抗肿瘤作用及其与顺铂的联合作用。经皮微粒介导法（基因枪系统）转染bax基因可抑制顺铂耐药的小鼠SCC的生长，并且bax基因与顺铂联合应用3天后对SCC的生长有明显的抑制作用（p<0.0001）。(2)本文研究了半胱天冬酶激活的脱氧核糖核酸酶（CAD）及其与顺铂（CDDP）联合应用的抗肿瘤作用。细胞凋亡过程中DNA的最终断裂受CAD的控制，基因枪介导的CAD基因对SCC的生长无明显抑制作用，但CAD与顺铂联合应用对SCC的生长有明显的抑制作用（p<0.05）。(3)研究了Fas-Fas配体系统的抗肿瘤作用。由于C3 H小鼠SCC不表达Fas配体，因此通过相同的基因枪将该配体递送到SCC中，并检查抗癌效果。结果表明，转染Fas配体的SCC细胞生长受到明显抑制（p<0.0003），存活时间明显延长，其中以bax基因+CDDP的效果最好。少

项目成果

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Nobuyuki Tanaka：“头颈癌中的 CDDP 耐药机制及其克服方法 - 重点关注 GS-X 泵增强药物流出和 DNA 修复能力”头颈肿瘤 25・1（1999）。

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