漆黄素及漆黄素纳米粒对卵巢癌细胞作用的研究

目的 探讨一种新的饮食性黄酮类化合物——漆黄素及漆黄素纳米粒对人卵巢癌细胞株SK0V3的抗增殖和促凋亡作用。 方法 体外实验采用MTT法测定漆黄素和漆黄素纳米粒作用后对卵巢癌细胞存活率的影响,选取下一步实验的药物浓度。裸鼠荷瘤后取肿瘤块制成SKOV3细胞混合液。取15只裸鼠腹腔接种人卵巢癌SKOV3细胞混合液肿瘤造模,第5天随机分成3组尾静脉注射药物,每3 d注射1次,每次0.2 mL,共12次:A组为肿瘤模型组（注射50 mL/L葡萄糖溶液）,B组为漆黄素组（注射1.25 mg/kg漆黄素）,C组为漆黄素纳米粒组（注射1.25 mg/kg漆黄素的米粒）。治疗结束后记录肿瘤质量,取心、肝、脾、肺和肾切片行HE染色观察组织病理损害,肿瘤组织切片通过Ki67免疫组织化学染色检测SKOV3细胞增殖,TUNEL法检测SKOV3细胞凋亡。 结果 MTT结果显示,随漆黄素及漆黄素纳米粒质量浓度的增加,SKOV3细胞的存活率减小;漆黄素及漆黄素纳米粒半数抑制浓度（IC₅₀）值分别为125~250 μg/mL和62.5~125 μg/mL,故选择工作浓度为125 μg/mL。共11只小鼠完成体内肿瘤造模并存活至实验结束（A组3只,B组4只,C组4只）。A组裸鼠腹腔内散在菜花状肿瘤结节,B、C组仅见极小颗粒状肿瘤结节,B、C组腹腔内肿瘤质量相当（P>0.05）,但均低于A组（P<0.05）。各组心、肝、脾、肺和肾切片HE染色均未见明显损害。与A组相比,B组和C组肿瘤组织增殖指数Ki67均降低,细胞凋亡率均升高（P<0.05）,但B组和C组之间差异无统计学意义（P>0.05）。 结论 漆黄素及漆黄素纳米粒均具有抗卵巢癌细胞增殖和促进其凋亡的作用,安全性好。

In vitro and in vivo Antitumor Effects of Fisetin and Fisetin Nanopartical on Ovarian Cancer

Objective To explore the effects of fisetin and fisetin nanopartical on anti-proliferation and apoptosis induction of human ovarian cancer cell line SKOV3. Methods The MTT assay was used to determine the survival rate of SKOV3 cell through apoptosis induction after fisetin and fisetin nanopartical treatment, and the drug concentration of the next experiment was selected. 15 nude mice vaccinated SKOV3 cells mixture mode randomly into three groups, treatment began on the fifth day, every 3 d injection times, each time 0.2 mL, a total of 12 times. Group A for the control (50 mL/L glucose solution), group B for fisetin (1.25 mg/kg), group C for fisetin nanopartical (1.25 mg/kg). After the treatment, tumor quality was recorded, and histopathological lesions were observed by HE staining on heart, liver, spleen, lung and kidney sections. The proliferation of SKOV3 cells was detected by Ki67 immunohistochemical staining in tumor tissue sections, and apoptosis of SKOV3 cells was detected by TUNEL assay. Results MTT test indicated that fisetin and fisetin nanopartical had inhibition effect on ovarian cancer cell in a dose-dependent manner. The half-maximal inhibitory concentration (IC₅₀) value for fisetin and fisetin nanopartical were 125-250 μg/mL and 62.5-125 μg/mL. Therefore, the working concentration was selected as 125 μg/mL. 11 mice were completed and survived until the end of the experiment (n=3 in group A, n=4 in group B, n=4 in group C). Group A nude mice abdominal cavity in cauliflower shape tumor nodules, B and C group only tiny granule tumor nodules, B and C group of intraperitoneal tumor mass is quite (P>0.05), but were lower than group A (P<0.05). No obvious damage was found in the hepatocytes, liver, spleen, lung, and kidney slices. Compared with group A, Ki67 index in group B and group C decreased, and cell apoptosis rate increased (P<0.05), but there was no significant difference between group B and group C (P>0.05).After treatment of fisetin and fisetin nanopartical, heart, liver, spleen, lung and kidney are safe without damage. Conclusion Fisetin and fisetin nanopartical have the effect of anti-ovarian cancer cells, and no organ damage has been found in both.