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目的:喜树碱(Camptothecin,CPT)是从植物喜树中分离得到的一种生物碱,在60年代就作为一种抗肿瘤药物用于临床。近年来,又从喜树中分离到比其更具抗痛活性的羟基喜树碱(HCPT),且无论从基础实验研究,抑或从临床应用上均发现HCPT是一种有效的抗癌药物。随着分子生物学理论和技术的不断发展,目前得知许多抗肿瘤药物是通过诱导肿瘤细胞凋亡来发挥其治疗作用的。为了阐明HCPT对大肠肿瘤细胞的杀伤作用是否同样经历着细胞凋亡的过程,我们选用TdT法进一步分析HCPT对人大肠癌细胞凋亡的影响。方法:1997年2月至1997年7月,取我院外科行手术切除的36例大肠癌标本,其中男17例,女19例,平均年龄(64.8±3.96)岁(29~82岁)。大肠痛标本中Dukes A期4例,B期20例,C_1期6例,C_2期1例,D期5例。Broder分级Ⅰ级1例,Ⅱ级34例,Ⅲ级1例。无菌情况下取手术切除的新鲜肿瘤组织约0.5g,获取单个肿瘤细胞后,向新鲜肿瘤细胞中分别加入羟基喜树碱0.1μg/ml,丹参(SM)6μI,5-FU10μg/ml,将加入药物的细胞置37℃5%C0_2中培养15h,取出肿瘤细胞采用TdT标记法(DNA断端标记法)检测调亡细胞。结果:36例人大肠肿瘤标本,分别加用不同药物进行细胞凋亡检测,羟基喜树碱的细胞凋亡率为29.2%,丹参为16.9%,HCPT+SM为38.6%,5-FU则为29 6%。同时对加用5-FU组?
Objective: Camptothecin (CPT) is an alkaloid isolated from the plant Camptotheca acuminatum. It was used as an anti-tumor drug in the clinic in the 1960s. In recent years, hydroxycamptothecin (HCPT) has been isolated from Camptotheca acuminata better than its anti-pain activity. HCPT is an effective anti-cancer drug from basic experimental studies and clinical applications. With the continuous development of molecular biology theory and technology, it is currently known that many antitumor drugs exert their therapeutic effects by inducing tumor cell apoptosis. To elucidate whether the killing effect of HCPT on colorectal cancer cells also undergoes apoptosis, we used TdT assay to further analyze the effect of HCPT on apoptosis of human colorectal cancer cells. METHODS: From February 1997 to July 1997, 36 patients with colorectal cancer underwent surgical resection in our hospital, including 17 males and 19 females, with an average age of (64.8±3.96) years (range, 29-82 years). In the large bowel pain specimens, 4 were in Dukes A stage, 20 in B stage, 6 in C 1 stage, 1 in C 2 stage, and 5 in D stage. Broder grading was grade I in 1 case, grade II in 34 cases, and grade III in 1 case. Under sterile conditions, about 0.5 g of fresh tumor tissue was removed from the tumor. After obtaining single tumor cells, hydroxycamptothecin 0.1 μg/ml, Salvia miltiorrhiza (SM) 6 μl, and 5-FU 10 μg/ml were added to fresh tumor cells. The drug-supplemented cells were cultured at 37°C in 5% C0_2 for 15 hours, and the tumor cells were taken out to detect apoptotic cells by TdT labeling (DNA fragmentation). RESULTS: Thirty-six human colorectal tumor specimens were treated with different drugs for apoptosis. The apoptosis rate of hydroxycamptothecin was 29.2%, Salvia miltiorrhiza was 16.9%, HCPT+SM was 38.6%, 5-FU was 29 6%. At the same time on the use of 5-FU group?