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

用ATP法评价139例头颈癌的化疗敏感率依次为：5-FU+GT；CDDP=CBDCA&GT；MTX+GT；PEP(23%)。因此，本研究的目的是克服化疗耐药与细胞凋亡相关的基因转导。1998年：在上述5种敏感药物中，顺铂被用于进一步的研究，因为顺铂是促进细胞凋亡的抗癌药物之一。CDDP诱导的细胞凋亡受谷胱甘肽和糖基化抑制剂衣霉素的调节。应用免疫组织化学方法对57例口咽鳞癌组织中Bax的表达与5年生存率的关系进行了研究。高表达促凋亡基因Bax的患者5年生存率明显提高。1999年：P53蛋白过度表达提示预后显著不良，而Bcl2蛋白过度表达与预后无关。…法检测P53、Bax、Bcl2蛋白表达与细胞凋亡率的相关性TUNEL染色越多，相关性不明显，而Bax阳性组的细胞凋亡率越高。针对临床病例高表达bax基因的患者预后较好的特点，研究了电穿孔转导bax基因的头颈部癌细胞系和耐药细胞系体外对顺铂的敏感性。转导bax基因的细胞株对CDDP的敏感性显著提高。表达bcl2基因的癌细胞株通过其反义基因显著提高了对CDDP的敏感性。2000：开展了促细胞凋亡基因化疗的体内研究。(1)研究促凋亡基因bax的抗癌作用及其与CDDP的联合作用。基因枪法经皮转移bax基因可抑制耐顺铂的小鼠鳞状细胞癌的生长，且bax基因与顺铂每隔3天联合应用可显著抑制小鼠鳞状细胞癌的生长(p&lt；0.0001)。(2)研究半胱氨酸氨基转移酶的抗癌作用及其与顺铂的联合作用。细胞凋亡过程中最终的DNA断裂是由CAD控制的，虽然基因枪单独携带的CAD基因经皮转移并未显示出明显的抑制作用，但联合应用CAD和CDDP可显著抑制SCC的生长(p&lt；0.05)。(3)研究了Fas-Fas配体系统的抗肿瘤作用。由于C3H小鼠SCC不表达Fas配体，因此用同一基因枪将Fas配体导入SCC，检测其抗肿瘤作用。转导Fas配体的鳞癌细胞生长受到明显抑制(p&lt；0.0003)，存活时间明显延长，其中bax基因+顺铂的抗肿瘤作用最为明显。较少

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