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

139例头颈部肿瘤的ATP法化疗敏感率依次为5-FU (27%) >CDDP=CBDCA>MTX>PEP（23%）。因此，本研究的目的是联合转染与细胞凋亡相关的基因来克服化疗耐药。1998：在上述5种敏感药物中，由于CDDP是促进细胞凋亡的抗癌药物之一，我们选用CDDP进行进一步的研究。CDDP引起的细胞凋亡受谷胱甘肽和tunicamycin（一种糖基化抑制剂）调控。应用免疫组织化学方法研究了57例口咽鳞状细胞癌中Bax表达与5年生存率的关系。表达高促凋亡基因bax的患者5年生存率明显提高。1999: p53过表达预后明显差，而Bcl-2过表达与预后无相关性。经TUNEL染色，p53、Bax、Bcl-2的表达与细胞凋亡率无显著相关性，但Bax阳性组细胞凋亡倾向较高。由于高表达bax基因的临床病例预后较好，因此研究电穿孔转染bax基因的头颈癌细胞株和抗CDDP细胞株是否在体外对CDDP更敏感。转染bax的细胞系对CDDP的敏感性显著提高。表达bcl-2的肿瘤细胞系通过其反义表达明显提高CDDP敏感性。2000年：用一些促进细胞凋亡的基因进行化疗的体内研究。(1)研究促凋亡基因bax的抗癌作用及其与CDDP的联合作用。bax基因经皮颗粒传递（基因枪系统）可抑制小鼠抗cddp SCC的生长。此外，bax基因与CDDP每隔3天联合使用可显著抑制小鼠SCC的生长（p<0.0001）。(2)研究了caspase-activated DNase的抗癌作用及其与CDDP的联合作用。凋亡过程中最终的DNA片段是由CAD控制的。虽然基因枪单独传递CAD基因经皮转移没有明显的抑制作用，但CAD和CDDP联合使用可显著抑制SCC的生长（p<0.05）。(3)研究了Fas-Fas配体体系的抗致癌作用。由于C3H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，并检测其抗癌作用。转染Fas配体的SCC生长受到显著抑制（p<0.0003），存活时间也显著延长。在上述3种促凋亡基因转染中，bax基因+CDDP的抗癌作用似乎最有效。少

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