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目的:观察夜香树甾体皂苷(SSCN)对肝癌HepG2移植瘤的抑制作用,并探讨其抑瘤的可能机制。方法:从夜香树叶中提取并鉴定SSCN;建立BALB/c裸鼠HepG2肝癌模型,建模成功后将荷人肝癌裸鼠随机分为5组:生理盐水组(NS)、沙利度胺组(TLD 200 mg·kg-1),SSCN低,中,高组(4,6,8 mg·kg-1,ip,隔日1次,连续30 d),观察SSCN的肿瘤抑制作用。治疗期间测量皮下移植瘤的长径和短径;30 d后处死裸鼠,剖取肿瘤,行常规制片,HE染色,镜下观察肿瘤病变;免疫组化染色,观察并检测移植瘤中增殖细胞核抗原PCNA和Ki-67蛋白的表达。结果:TLD组裸鼠ip对肝癌HepG2 BALB/c裸鼠相对肿瘤增殖率为42.19%,SSCN 4,6,8 mg·kg-1的增殖率分别为48.26%,42.94%,41.57%,随着剂量的增高其相对肿瘤增殖率逐渐下降,SSCN抑制肿瘤生长的作用存在剂量依赖性。镜下观察,SSCN高剂量组见到大量坏死细胞外,部分肿瘤细胞体积较小,细胞核浓染。NS组荷瘤小鼠的肿瘤细胞中PCNA和Ki-67蛋白的表达率明显增高;而与NS组比较,TLD组和SSCN高剂量组PCNA和Ki-67蛋白的表达显著下降(P<0.05)。结论:夜香树甾体皂苷体内对BALB/c小鼠肝癌移植瘤有一定的抑制作用,其作用机制可能是通过下调PCNA和Ki-67基因蛋白的表达,从而抑制肿瘤细胞的增殖。
OBJECTIVE: To observe the inhibitory effect of SSCN on hepatocellular carcinoma HepG2 xenografts and to explore the possible mechanism of antitumor activity. Methods: SSCN was extracted and identified from the leaves of C. sinensis. HepG2 hepatocellular carcinoma model was established in BALB / c nude mice. After successful modeling, nude mice bearing human hepatocellular carcinoma were randomly divided into 5 groups: normal saline group (NS), thalidomide group TLD 200 mg · kg-1), SSCN low, middle and high group (4, 6, 8 mg · kg-1, ip, every other day for 30 days). The long and short diameters of the subcutaneously transplanted tumors were measured during the treatment. After 30 days, the nude mice were sacrificed and the tumors were dissected. The tumors were routinely prepared and stained with hematoxylin and eosin (HE). The tumor lesions were observed under the microscope. The expression of nuclear antigen PCNA and Ki-67 protein. Results: The relative tumor proliferation rate of nude mice in TLD group was 42.19% for HepG2 BALB / c nude mice and 48.26%, 42.94% and 41.57% for SSCN 4, 6 and 8 mg · kg-1, respectively The dose-dependent increase of its relative tumor growth rate decreased, and the effect of SSCN on tumor growth was dose-dependent. Microscope observation, high-dose SSCN group to see a large number of necrotic cells, some tumor cells smaller, nuclear staining. Compared with NS group, the expression of PCNA and Ki-67 protein in tumor cells of NS group were significantly decreased (P <0.05) . Conclusion: The sterilized saponin of Syringa obscurus can inhibit the transplanted hepatocellular carcinoma of BALB / c mice in vitro. The mechanism may be that it inhibits the proliferation of tumor cells by down-regulating the expression of PCNA and Ki-67 gene protein.