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目的:探讨H-ras12V转基因小鼠的肝癌发生与遗传不稳定性的关系,以及活性氧(reactive oxygen species,ROS)在其中的作用。方法:用微卫星标志物的PCR-放射自显影法检测H-ras12V转基因雌雄小鼠肝癌及其周围组织的微卫星杂合性缺失(loss of heterozygosity,LOH)、杂合性保留(retention of heterozygosity,ROH)和微卫星不稳定性(microsatellite instability,MSI)。用实时荧光定量PCR和FCM法分别分析该转基因雌雄小鼠肝组织中H-ras12V癌基因表达水平和ROS水平。结果:仅雌雄转基因小鼠肝癌组织中伴有LOH发生;所有小鼠均发现伴有ROH,但转基因小鼠肝癌组织中ROH检出率与非转基因小鼠肝组织相比显著增高(P<0.05);所有组织标本中均未检测出MSI。转基因小鼠的H-ras12V癌基因表达和ROS水平均显著高于非转基因小鼠,且雄性小鼠显著高于雌性小鼠(P<0.05)。结论:H-ras12V癌基因可能通过诱导ROS水平增高,进一步引起遗传不稳定性。这可能是H-ras12V转基因小鼠肝癌发生的一条重要途径。
Objective: To investigate the relationship between the occurrence of hepatocellular carcinoma (HCC) and genetic instability in H-ras12V transgenic mice and the role of reactive oxygen species (ROS) in H-ras12V transgenic mice. METHODS: The microsatellite heterozygosity (LOH) and retention of heterozygosity (HOH) of H-ras12V transgenic male and female mice hepatocarcinoma and their surrounding tissues were detected by PCR-autoradiography. , ROH) and microsatellite instability (MSI). The expression of H-ras12V oncogene and the level of ROS in liver tissue of the transgenic male and female mice were analyzed by real-time fluorescence quantitative PCR and FCM respectively. RESULTS: Only LOH occurred in HCC tissues of both male and female transgenic mice. ROH was detected in all mice, but the detection rate of ROH in transgenic mice was significantly higher than that in non-transgenic mice (P <0.05) ); MSI was not detected in all tissue samples. The H-ras12V oncogene expression and ROS level in transgenic mice were significantly higher than those in non-transgenic mice, and the male mice were significantly higher than female mice (P <0.05). Conclusion: The H-ras12V oncogene may further induce genetic instability by inducing the increase of ROS level. This may be H-ras12V transgenic mouse liver cancer an important way.