肺岩宁颗粒干预细胞周期G1/S检测点调控信号抗Lewis肺癌细胞增殖的研究

目的 观察肺岩宁颗粒对Lewis肺癌小鼠肿瘤生长、肿瘤细胞周期分布，以及G1/S检测点调控信号RB－E2F1生物轴的影响。 方法 采用C57BL/6小鼠，造模并随机分为6组：模型组、顺铂组、肺岩宁煎剂组（煎剂组）、肺岩宁颗粒低剂量组（低剂量组，0.3 g）、肺岩宁颗粒中剂量组（中剂量组,0.6 g）、肺岩宁颗粒高剂量组（高剂量组,1.2 g）。造模后顺铂组1、3、5天腹腔注射顺铂0.1 mg,其他各组分别灌胃给药14天，观察各组治疗后抑瘤率、瘤重、体重，流式细胞术检测细胞周期时相分布，RT-PCR测定肿瘤组织视网膜母细胞瘤蛋白（RB）－腺病毒E2启动子结合转录因子（E2F1）的mRNA表达,Western blot检测RB、E2F1蛋白表达。 结果 各用药组瘤重低于模型组（P<0.05，P<0.01），中剂量组体重明显高于模型组和顺铂组（P<0.05，P<0.01）。流式细胞术发现中剂量组的G0/G1比例较模型组、顺铂组和煎剂组显著增多（P<0.01），癌细胞增殖指数明显降低（P<0.01，P<0.05）。RT-PCR显示中剂量组E2F1 mRNA水平明显低于模型组、顺铂组和煎剂组(P<0.01)；中剂量组RB mRNA表达明显高于模型组、顺铂组和煎剂组 (P<0.01)。Western blot分析显示中剂量组较模型组和顺铂组明显抑制E2F1蛋白表达(P<0.05)，并较模型组、顺铂组和煎剂组明显增加RB蛋白表达(P<0.05)。 结论 肺岩宁颗粒抑制Lewis肺癌肿瘤生长与干预细胞周期时相分布有关，能使癌细胞较多的阻滞在细胞周期G0/G1期，通过抑制E2F1，增强RB表达，从而干预细胞周期G1/S检测点信号通路中RB－E2F1生物轴实现抗肺癌增殖。

Effect of Feiyanning Granule in Antagonizing Lewis Lung Cancer Cell Proliferation through Cell Cycle G1/S Checkpoint Dominating Signaling Intervention

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 dl, 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- E2FlmRNA 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 （P1） 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）. Western blot analysis showed the protein expression of E2F1 was lower in clrout） 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.