槲寄生碱抑制肺癌细胞增殖作用初步观察

Inhibitory effects of mistletoe alkali on human lung adenocarcinoma SPC-A1and squamous SK-MES-1cells

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 alkali on human lung adenocarcinoma carcinoma cell line SPC-A1 and lung squamous carcinoma cell line SK-MES-1 in vitro. METHODS：The inhibitory effect on the cells treated by mistletoe alkali with different concentration was evaluated by methyl thiazolyl tetrazolium （MTT） assay for 24 and 48 hours respectively. The cells treated by 5-fluorouracil （5-FU） were used as positive control group,while the cells without any treatment as negative control group. Different doses of mistletoe alkali were used as experimental group. The 50% inhibiting concentration （ICs0） was assessed by cell growth curve. The rate of apoptosis for 24h was detected by Flow Cytometry. RESULTS： Mistletoe alkali did exert inhibitory effect on growth of cells in dose and time dependent man- ner. ICs0 of mistletoe alkali to SPC-A1 and SK-MES-1 for 24 h was （14.43±0.53） μg/mL and （8.09±0.40） μg/mL re- spectivelyICs0 was （12.11±0.25） μg/mL and （6.43±0.33） μg/mL respectively for 48 h. ICs0 of 5-FU to SPC-A1 and SK-MES-1 for 24 h were （4. 79±0. 45） μg/mL and （5. 15±0. 23） μg/mL respectively; for 48 h were （4. 35± 0.41） /μg/mL and （4. 11 ± 0. 38） μg/mL respectively; Apoptosis rate was （0.43 ± 0.01） in control group, （31.09 ±0.05） M for SPC-A1 cell line in high dose group, the concentration of mistletoe alkali was 28.86 /μg/mL, and （18.19 ± 0.02） 0/oo in middle dose group,the concentration of mistletoe alkali was 14.43 /μg/mL, （7.99±0.05） % in low dose group, the concentration of mistletoe alkali was 7.22 μg/mL. While for SK-MES-1 cell line, the apoptosis rate was （38.24±0.03） % in high dose group, the concentration of mistletoe alkali was 16.18 μg/mL, and （21.81± 0.01）% in middle dose group, the concentration of mistletoe alkali was 8.09 /μg/mL, （9.11±0.01） % in low dose group, the concentration of mis tletoe alkali was 4.05 μg/mL. Compared with that of negative control group, mistletoe alkali led to dramatic increase in ap-optosis rate of SPC-A1 and SK-MES-1 cell line （P〈0.05）, which was growing as the concentration increasing （P〈 0.05）. CONCLUSIONS：Mistletoe alkali can inhibit the proliferation and induce apoptosis of SPC-A1 and SK-MES-1 cell line obviously, which provides an experimental reference for application of mistletoe alkali to treatment of human lung car cinoma.