【作者】 钱军；

【导师】 刘沈林；

【作者基本信息】 南京中医药大学 ， 中西医结合临床， 2009， 博士

【摘要】 原发性肝癌是我国最常见的恶性肿瘤之一,严重威胁人类的健康和生命。由于该病起病隐袭、进展迅速、早期诊断率低,大多数病人在确诊时已达中晚期,往往失去了手术治疗的机会,只能进行姑息性的系统治疗和支持对症治疗。以往认为肝癌化疗效果差,易产生多药耐药,有效率低,加上化疗药物毒副作用较大,有关研究进展缓慢,急需突破。研究新的高效低毒的治疗药物和合理的中西医结合治疗方案以提高肝癌的治疗效果是肿瘤工作者探索并亟待攻克的难题。消癌平注射液系由植物通关藤的根、茎或叶经现代工艺所制成的中药制剂,为国家正式上市产品。临床被应用于治疗各种癌症和咳嗽、哮喘等疾病。但目前基础研究十分薄弱,临床研究相当粗浅,有必要对消癌平注射液抗肝癌及其作用机制进行深入而系统的研究。通过文献考证通关藤制剂的抗肿瘤作用及其机制,明确了通关藤制剂的植物来源和抗肿瘤有效成分,为该类中药有效成分抗肿瘤研究提供了理论依据,从而更好地对其开发利用。实验研究表明:消癌平注射液对体内外肝癌细胞均有一定的抑制作用,其抑制作用随着时间的延长和浓度的增加而逐渐增强。消癌平的抗癌作用可能具有多样化和多靶点的特点,能使癌细胞G₀/G₁期细胞增多,G₂/M期细胞减少,并在形态学上出现诱导凋亡的一些表现;可以抑制Ki-67蛋白表达,使得凋亡抑制蛋白Bcl-2表达减弱,促凋亡蛋白Bax表达增强。顺铂是原发性肝癌的基本化疗药物之一,但是毒性显著,应用明显受限;如今多种新型铂类药物已经陆续问世,高效低毒,明显提高了其他消化道癌的化疗效果且耐受性良好。在体外实验研究中,比较了不同的铂类药物对于肝癌细胞株HEPG2、SMMC-7721和Bel-7402的抑制作用,为临床上选择恰当的铂类药物治疗肝癌提供参考依据。采用MTT法检测加入不同铂类药物后对于肝癌细胞的抑制作用,观察给药前、后肝癌细胞的形态学变化,计算半数抑制浓度,绘制肝癌细胞生长曲线。流式细胞分析给药后细胞周期的变化。结果显示给予铂类药物后肝癌细胞生长减慢,胞浆内颗粒增多;随着给药时间延长,对肝癌细胞的抑制作用逐渐增强。对于肝癌细胞的抑制作用,顺铂和奥沙利铂强于洛铂、奈达铂和卡铂。流式细胞分析不同铂类药物对细胞周期的变化趋于一致。因此,在体外研究中明确了铂类药物对于肝癌细胞具有抑制作用,顺铂仍为基本用药,而奥沙利铂具有一定的优势,可推荐在临床上用于治疗原发性肝癌。联合作用的实验研究表明,消癌平与铂类药物对肝癌细胞的抑制作用具有协同作用,这种协同作用需在消癌平达到一定的剂量水平时表现明显;可以增强铂类药物对肝癌的抑制作用。对其协同作用的机制尚待深入研究。更多还原

【Abstract】 Primary liver cancer is one of the most common malignant tumor in China, a serious threat to human health and life. Because of hidden hit of the disease onset, progress rapidly, the low rate of early diagnosis, most patients had reached advanced stage of disease at the time of diagnosis. Surgical treatment is often lost opportunity with these patients and those can only palliative treatment of the symptomatic treatment and support. Consider the effect of chemotherapy was bad. easy to produce multi-drug resistance, have low efficiency, coupled with drug side effects of chemotherapy drugs, the slow progress of the study, much-needed breakthrough. Research on new drugs and efficient treatment of low toxicity and reasonable treatment in traditional Chinese and western medicine in order to improve the treatment of liver tumor are need to explore and overcome problems.Xiaoaiping Injection came from the rattan plant root, stem or leaf and was made by the modern process of Marsdenia tenacissima. It being used in clinical treatment of various types of cancer and cough, asthma and other diseases. But it was very weak in basic research and clinical research rather shallow, there have a requirement with depth and systematic study for Xiaoaiping injection and its mechanism of anti-liver cancer.The article reviewed the progress in the research of antitumor activities and mechanisms of Marsdenia tenacissima. Its origin producing area and antitumor constituents were analyzed. It provides theoretical evidences for study on constituents in this traditional Chinese medicine. Futher studies on the species in Marsdenia tenacissima are needed for better medicinal utilization. Experimental results showed that: Xiaoaiping injection inhibited liver cancer cells in vitro and in vivo, its inhibition increased with the time and concentration gradually. Xiaoaiping had anticancer effects of diversification and target many of the characteristics of cancer cells. It could increase cells in G₀/G₁ phase, decrease cells in G₂/M phase and morphological appeared on some of the performance of induction of apoptosis; It could inhibit the expression of Ki-67 protein, the inhibitor of apoptosis protein Bcl-2 expression decreased, Bax protein expression enhanced.Cisplatin was one of the essential drag in chemotherapy of hepatocellular carcinoma. It was restricted in clinical application because of its toxicity. To explore the different effect of different platinum complexes in human hepatocellular carcinoma cell line （HEPG2, SMMC-7721 and Bel-7402） by experiment in vitro, this experiment intent to dedicate evidence for choosing platinum complexes in clinical chemotherapy of hepatocellular carcinoma. To observe the cellular morphology after being given different platinum complexes. To calculate inhibitory concentration （IC50） and get cell growth curve by MTT. The cell cycle was analyzed by flow cytometry. We observed the decrease of cell growth, increase of particle of endochylema; increase of the depressant effect to human hepatocellular carcinoma cell line with the increase of time being given platinum complexes. Cisplatin and Oxaliplatin were better than lobaplatin, nedaplatin and carboplatin in the depressant effect to human hepatocellular carcinoma cell line. FCM analysis showed that there appeared similar change in cell cycle after being given platinum complexes. Those showed platinum complexes had depressant effect to human hepatocellular carcinoma cell line in vitro. Cisplatin was the essential drug and Oxaliplatin was another better choice for clinical chemotherapy of hepatocellular carcinoma. The two drags were prefered in chemotherapy of hepatocellular carcinoma.The role of the United experimental studies had shown that Xiaoaiping with platinum-based drags on liver cancer cells in vitro displayed synergy. Xiaoaiping needed to reach a certain dose level with the synergy of performance significantly. It could enhance platinum-based drags on inhibition of liver cancer. The mechanism of their synergy had yet to be studied in future.更多还原