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应用四甲基偶氮唑盐(MTT)比色法进行大肠癌细胞体外培养的化疗药敏试验,结合 PCR-SSCP 银染技术分析大肠癌 p53基因突变,结果显示可检测到的20例大肠癌细胞对5-氟脲嘧啶(5-Fu)最为敏感(IR=54.7±14.1%),丝裂霉素(MMC)(IR=43.0±1.14%,P<0.05)和阿霉素(ADM)(IR=42.0±11.1%,P<0.05)次之,对顺铂(DDP)不敏感(IR=25.9±12.8%,P<0.01)。不同的化疗药敏组间存在显著性差异,P<0.001。p53基因突变率为45%(9/20例)。点相关分析结果证实 MMC 和 ADM 的药敏结果与 p53基因状态显著正相关(Rmmc=0.589,Radm=0.628,P<0.05),其所诱导的大肠癌细胞凋亡为 p53基因依赖性的。而5-Fu 和 DDP 所诱导的大肠癌细胞凋亡则为非 p53基因依赖性的,提示可能有其他如 c-myc、c-fos、Bcl-2或 ras 基因在5-Fu 和 DDP 所诱导的大肠癌细胞凋亡中起着主要的调控作用。
MTT colorimetric method was used to perform chemotherapeutic susceptibility testing of colorectal cancer cells in vitro. PCR-SSCP silver staining technique was used to analyze colorectal cancer p53 mutations. The results showed that 20 colorectal cancers could be detected. The cells were most sensitive to 5-fluorouracil (5-Fu) (IR=54.7±14.1%), Mitomycin (MMC) (IR=43.0±1.14%, P<0.05) and Adriamycin (ADM) ( IR = 42.0±11.1%, P<0.05) followed by cisplatin (DDP) insensitivity (IR=25.9±12.8%, P<0.01). There was a significant difference between different chemotherapeutic groups, P<0.001. The mutation rate of p53 gene was 45% (9/20 cases). The results of point correlation analysis confirmed that the drug sensitivity of MMC and ADM was positively correlated with the p53 gene status (Rmmc=0.589, Radm=0.628, P<0.05). The apoptosis of colorectal cancer cells induced by p53 gene was dependent on p53 gene. Apoptosis of colorectal cancer cells induced by 5-Fu and DDP was non-p53-dependent, suggesting that other genes such as c-myc, c-fos, Bcl-2, or ras may be induced by 5-Fu and DDP. The colorectal cancer cell apoptosis plays a major regulatory role.