[肺岩宁颗粒通过细胞周期G1/S检查点主导信号干预拮抗Lewis肺癌细胞增殖的作用]

[Effect of felyanning granule in antagonizing Lewis lung cancer cell proliferation through cell cycle G1/S checkpoint dominating signaling intervention].

作者信息

Zheng Zhan, Wang Ju-yong, Xu Zhen-ye

机构信息

Long-hua Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2009 Nov;29(11):1018-22.

PMID:20329615

Abstract

OBJECTIVE

To observe the effect of Feiyanning Granule (FYN) on tumor growth and cell cycle distribution in mice with Lewis lung cancer, as well as its influence on G1/S cell cycle checkpoint dominating signaling RB-E2F1 bio-axis.

METHODS

Modeled C57BL/6 mice were randomly divided into 6 groups: the model group (A), the DDP treated group (B) peritoneally injected with cisplatin 0.1 mg on d1, d3 and d5 after modeling, and the 4 FYN treated groups (C-F), administered via gastrogavage with FYN Decoction, and FYN Granule in small-, median- and high- dose respectively for 14 days. The tumour inhibiting rate, tumour weight, and body weight of mice were observed after treatment; cell cycle distribution was detected by flow cytometry (FCM), RB-E2F1mRNA expressions in tumour tissue were analyzed by RT-PCR, and their protein expressions by Western blot.

RESULTS

Tumour weight in the 5 treated groups was lower than that in the model group (P < 0.05, P < 0.01). Body weight in group E was significantly higher than that in group A and B (P < 0.05, P < 0.01). FCM analysis showed the proportion of G0/G1 phase was higher in group E than in group A, B and C (P < 0.01), and cancer cell proliferation index (PI) in group E was lower than in group B (P < 0.05, P < 0.01). RT-PCR showed mRNA level of E2F1 was lower, but that of RB was significantly higher in group E than those in group A, B and C respectively (P < 0.01). Westem blot analysis showed the protein expression of E2F1 was lower in group E and B than that in group A (P < 0.05), while the protein expression of Rb in group E was higher than that in group A, B and C (P < 0.05).

CONCLUSION

The effect of FYN in inhibiting Lewis lung cancer growth was related to its intervention on cancer cell cycle distribution which blocks most tumor cells in G0/G1 phase. Moreover, FYN can reduce MDM2 expression, enhance P53 expression to influence cell cycle G1/S checkpoint dominating signaling, so as to achieve the effect of antagonizing lung cancer cell proliferation.

摘要

目的

观察肺岩宁颗粒（FYN）对Lewis肺癌小鼠肿瘤生长及细胞周期分布的影响，以及对G1/S细胞周期关卡主导信号RB-E2F1生物轴的影响。

方法

将造模后的C57BL/6小鼠随机分为6组：模型组（A）；顺铂治疗组（B），造模后第1、3、5天腹腔注射顺铂0.1mg；4个FYN治疗组（C-F），分别灌胃给予肺岩宁水煎剂、小剂量、中剂量和高剂量肺岩宁颗粒，连续给药14天。观察治疗后小鼠的抑瘤率、肿瘤重量及体重；采用流式细胞术（FCM）检测细胞周期分布，采用RT-PCR分析肿瘤组织中RB-E2F1mRNA表达，采用蛋白质免疫印迹法分析其蛋白表达。

结果

5个治疗组的肿瘤重量均低于模型组（P<0.05，P<0.01）。E组小鼠体重显著高于A组和B组（P<0.05，P<0.01）。FCM分析显示，E组G0/G1期比例高于A组、B组和C组（P<0.01），E组癌细胞增殖指数（PI）低于B组（P<0.05，P<0.01）。RT-PCR显示，E组E2F1mRNA水平低于A组、B组和C组，而RB mRNA水平显著高于A组、B组和C组（P<0.01）。蛋白质免疫印迹分析显示，E组和B组E2F1蛋白表达低于A组（P<0.05），而E组Rb蛋白表达高于A组、B组和C组（P<0.05）。