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目的:研究槲寄生碱对人肺腺癌细胞株SPC-A1及肺鳞癌细胞株SK-MES-1的抑制作用。方法:采用四甲基噻唑氮蓝(methyl thiazolyl tetrazolium,MTT)比色法检测不同浓度槲寄生碱作用24和48h对细胞增殖的影响,以含不同浓度的5-氟尿嘧啶(5-FU)细胞组为阳性对照,不含药物的细胞组为阴性对照,计算半数抑制浓度(IC50)。流式细胞术(flow cytometry,FCM)检测高、中和低3组浓度的槲寄生碱作用24h后的细胞的凋亡率。结果:不同浓度槲寄生碱作用24h,对SPC-A1细胞的IC50为(14.43±0.53)μg/mL,对SK-MES-1细胞的IC50为(8.09±0.40)μg/mL。其作用48h,对SPC-A1细胞的IC50为(12.11±0.25)μg/mL,对SK-MES-1细胞的IC50为(6.43±0.33)μg/mL。并且抑制作用随碱浓度的升高和作用时间的延长而增加,P<0.05。阳性对照组5-FU作用24h,对SPC-A1细胞的IC50为(4.79±0.45)μg/mL,对SK-MES-1细胞的IC50为(5.15±0.23)μg/mL。作用48h,对SPC-A1细胞的IC50为(4.35±0.41)μg/mL,对SK-MES-1细胞的IC50为(4.11±0.38)μg/mL。FCM检测SPC-A1细胞未加入药物的对照组凋亡率为(0.43±0.01)%,高剂量组药物浓度28.86μg/mL,凋亡率为(31.09±0.05)%,中剂量组药物浓度14.43μg/mL,凋亡率为(18.19±0.02)%,低剂量组药物浓度7.22μg/mL,凋亡率为(7.99±0.01)%。SK-MES-1细胞未加入药物的对照组凋亡率为(0.57±0.02)%,高剂量组药物浓度16.18μg/mL,凋亡率为(38.24±0.03)%,中剂量组药物浓度为8.09μg/mL,凋亡率为(21.81±0.01)%,低剂量组药物浓度4.05μg/mL,凋亡率为(9.11±0.01)%,与没有加入槲寄生碱的细胞相比,加入槲寄生碱后SPC-A1细胞与SK-MES-1细胞凋亡率均明显增加,P<0.05,并且凋亡率随槲寄生碱剂量的增加而升高,P<0.05。结论:槲寄生碱能够抑制人肺腺癌细胞株SPC-A1及肺鳞癌细胞株SK-MES-1的增殖,并能促进细胞的凋亡,为槲寄生碱应用于肺癌的临床治疗提供了实验依据。
Objective: To study the inhibitory effect of mistletoe on human lung adenocarcinoma cell line SPC-A1 and lung squamous carcinoma cell line SK-MES-1. Methods: MTT assay was used to detect the effects of different concentration of mistletoe alkaloids on cell proliferation at 24 and 48 hours. MTT assay was used to detect cell proliferation in vitro. For the positive control, drug-free cell group negative control, calculated half-value inhibition concentration (IC50). Flow cytometry (flow cytometry, FCM) detection of high, medium and low concentrations of three groups of mistletoe alkali after 24h apoptosis rate. Results: The IC50 of SPC-A1 cells was (14.43 ± 0.53) μg / mL and the IC50 of SK-MES-1 cells was (8.09 ± 0.40) μg / mL for 24 h. The IC50 was (12.11 ± 0.25) μg / mL for SPC-A1 cells and (6.43 ± 0.33) μg / mL for SK-MES-1 cells for 48h. And the inhibition increased with the increase of alkali concentration and prolongation of action time, P <0.05. The IC50 of positive control group was (4.79 ± 0.45) μg / mL for SPC-A1 cells and (5.15 ± 0.23) μg / mL for SK-MES-1 cells for 24h. After 48 hours, the IC50 of SPC-A1 cells was (4.35 ± 0.41) μg / mL and that of SK-MES-1 cells was (4.11 ± 0.38) μg / mL. FCM showed that the apoptosis rate of the control group without SPC-A1 cells was (0.43 ± 0.01)%, the high dose group was 28.86μg / mL, the apoptosis rate was (31.09 ± 0.05)%, the middle dose group was 14.43 μg / mL, the apoptosis rate was (18.19 ± 0.02)% and the low dose group was 7.22 μg / mL. The apoptosis rate was (7.99 ± 0.01)%. The apoptotic rate of SK-MES-1 cells was (0.57 ± 0.02)% in control group, 16.18μg / mL in high dose group, and (38.24 ± 0.03)% in SK-MES- 8.09μg / mL, the apoptosis rate was (21.81 ± 0.01)%, the low dose group was 4.05μg / mL and the apoptosis rate was (9.11 ± 0.01)%. Compared with the cells without the addition of mistletoe, The apoptosis rate of SPC-A1 cells and SK-MES-1 cells were significantly increased after parasitism (P <0.05), and the apoptosis rate increased with the increase of alkali mistletoe dosage, P <0.05. Conclusions: Mistletoe alkaloids can inhibit the proliferation of human lung adenocarcinoma cell line SPC-A1 and lung squamous cell carcinoma cell line SK-MES-1, and promote the apoptosis of cells, providing the clinical application of mistletoe alkaloids in lung cancer Experimental basis.