芹菜素影响非小细胞肺癌A549细胞顺铂敏感性的RAD51基因调控机制研究 点击下载
| 论文标题： | 芹菜素影响非小细胞肺癌A549细胞顺铂敏感性的RAD51基因调控机制研究 |
| 英文标题： | |
| 中文摘要： | 目的：从RAD51基因途径研究芹菜素对非小细胞肺癌（NSCLC）A549细胞顺铂敏感性的影响及其机制。方法：取人肺癌顺铂耐药细胞A549/DDP，分为对照组（空白培养基）、顺铂组（5g/L）和芹菜素低、高剂量组（10、20μmol/L），采用MTT法检测A549/DDP细胞生长，采用AnnexinⅤ/PI双染色法结合流式细胞术检测其凋亡情况。取A549/DDP细胞，分为顺铂单用组（1、2、4、8、16μg/mL）和芹菜素联用组（10μmol/L，在顺铂基础上加用），采用MTT法测定并计算细胞增殖抑制率；采用回归分析模型计算药物的半数抑制浓度（IC50），并以此计算芹菜素的逆转指数。将18只裸鼠随机分为对照组、顺铂单用组和芹菜素联用组，每组6只，接种A549/DDP细胞使形成移植瘤后，分别腹腔注射生理盐水、顺铂（2mg/kg，隔天给药1次）、顺铂（剂量、用法同前）+芹菜素药液（30mg/kg，每天给药1次），连续给药18d，测量小鼠的体质量和移植瘤质量并计算抑瘤率。取人肺癌细胞A549和A549/DDP，分为正常组（A549细胞）、对照组（A549/DDP细胞）、顺铂组（5g/L，A549/DDP细胞）和芹菜素低、高剂量组（10、20μmol/L，A549/DDP细胞），分别采用实时荧光定量聚合酶链反应和Westernblotting法检测细胞中RAD51的mRNA及其蛋白表达情况。结果：与对照组比较，芹菜素低、高剂量组细胞增长数均显著降低，凋亡率均显著升高且显著高于顺铂组（P＜0.05或P＜0.01）。联用芹菜素后，A549/DDP细胞的增殖抑制率均较相应质量浓度的顺铂单用组显著升高（P＜0.05）；芹菜素联用组的IC50为（5.81±0.47）μg/mL，显著低于顺铂单用组的IC50（14.44±0.52）μg/mL（P＜0.05）；逆转指数为2.49。裸鼠抑瘤实验结果显示，联用芹菜素后，A549/DDP荷瘤裸鼠抑瘤率为68.72％，显著低于顺铂单用组的33.82％（P＜0.05）。与正常组A549细胞比较，对照组A549/DDP细胞中RAD51mRNA及其蛋白的相对表达量均显著升高（P＜0.05）；与对照组A549/DDP细胞比较，芹菜素低、高剂量组A549/DDP细胞中RAD51mRNA及其蛋白的相对表达量均显著降低（P＜0.05）；与顺铂组A549/DDP细胞比较，芹菜素低、高剂量组A549/DDP细胞中RAD51mRNA及其蛋白的相对表达量均显著降低（P＜0.05）。结论：芹菜素能够有效逆转人肺癌顺铂耐药细胞A549/DDP的耐药性，其机制可能与下调RAD51基因转录及其蛋白表达有关。 |
| 英文摘要： | OBJECTIVE：To study the effects and mechanism o f apigenin on cisplatin sensitivity of NSCLC A 549 cells by regulating RAD51 gene. METHODS ：Human lung cancer cisplatin-resistant cells A 549/DDP were selected and divided into control group（blank culture medium ），cisplatin group （5 g/L），apigenin low-dose and high-dose groups （10，20 μmol/L）. MTT assay was used to detect the growth of A 549/DDP cells ，while the Annexin Ⅴ/PI double staining combined with flow cytometry were used to detect the apoptosis. A 549/DDP cells were collected and divided into cisplatin alone group （1，2，4，8，16 μg/mL）， apigenin combination group （10 μmol/L，based on cisplatin ）. MTT method was used to determine and calculate inhibitory rate of cell proliferation. IC 50 values of drugs were calculated by regression model ，and reversion index of apigenin was calculated. 18 nude mice were randomly divided into control group ，cisplatin alone group and apigenin combination group ，with 6 mice in each group. After A 549/DDP cells were inoculated to form the transplanted tumor ，normal saline ，cisplatin（2 mg/kg，once every other day ）， cisplatin（the same dosage and usage ）+apigenin solution （30 mg/kg，once a day ）were injected intraperitoneally respectively. After 18 days of continuous administration ，the body weight of mice and the mass of the transplanted tumor were detected and the tumor inhibition rate was calculated. Human lung cancer cells A 549 and A 549/DDP were collected and divided into normal group （A549 cells），control group （A549/DDP cells ），cisplatin group （5 g/L，A549/DDP cells ）and apigenin low-dose and high-dose groups （10，20 μmol/L，A549/DDP cells ），respectively. mRNA and protein expression of RAD51 were detected by real-time fluorescence quantitative PCR and Western blotting assay. RESULTS ： Δ 基金项目：四川省教育厅（自筹经费）科研项目（No.12ZB227） ＊讲师，硕士。研究方向：肿瘤病理。E-mail：molin212@163.com Compared with control group ，the cell growth of apigenin # 通信作者：副教授，硕士生导师，硕士。研究方向：肿瘤病理。 low-dose and high-dose groups were decreased significantly ， E-mail：xinrongH292@163.com apoptosis rates of them w ere increased significantly and higher ·708· China Pharmacy 2020Vol. 31 No. 6 中国药房 2020年第31卷第6期 than those of cisplat in group （P＜0.05 or P＜0.01）. After combined with apigenin ，proliferation inhibition rate of A 549/DDP cells was increased significantly ，compared with cisplatin alone group with the same concentration （P＜0.05）. The IC 50 in the apigenin combination group was （5.81±0.47）μg/mL，significantly lower than （14.44±0.52）μg/mL in cisplatin alone group（P＜0.05），and reversal index of apigenin was 2.49. The results of nude mice tumor inhibition experiment showed that after combined with apigenin，tumor inhibition rate of A 549/DDP bearing nude mice was 68.72％，significantly lower than 33.82％ in cisplatin alone group. Compared with A 549 cells of normal group ，relative expression of RAD51 mRNA and protein were increased significantly in A549/DDP cells of control group （P＜0.05）. Compared with A 549/DDP cells of control group ，relative expression of RAD51 mRNA and protein in A 549/DDP cells were decreased significantly in apigenin low-dose and high-dose groups （P＜0.05）. Compared with cisplatin group ，relative expression of RAD51 mRNA and protein in A 549/DDP cells of apigenin low-dose and high-dose group were decreased significantly （P＜0.05）. CONCLUSIONS ：Apigenin can effectively reverse drug resistance of cisplatin-resistant A 549/DDP cells in human lung cancer. The mechanism may be related to the reduction of RAD51 gene transcription and protein expression. |
| 期刊： | 2020年第31卷第06期 |
| 作者： | 莫琳，刘馨，杨慧敏，何欣蓉，王小林，唐春红 |
| 英文作者： | MO Lin，LIU Xin，YANG Huimin ，HE Xinrong ，WANG Xiaolin ，TANG Chunhong |
| 关键字： | 芹菜素；RAD51；非小细胞肺癌；顺铂；耐药；敏感性 |
| KEYWORDS： | Apigenin；RAD51；Non-small c ell lung cancer ；Cisplatin；Drug resistance ；Sensitivity |
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