Enhancement of radiosensitivity of lung adenocarcinoma using a decoction from the Fuzhengzengxiao formula

OBJECTIVE: To study the effects of a decoction of Fuzhengzengxiao formula on lung adenocarcinoma regarding the inflammatory protein S100A9 known to enhance cancer cell sensitivity.METHODS: A nude mouse model of human lung adenocarcinoma was established. The mice were randomly divided into four groups using the random number table method: Group Ⅰ, control;Group Ⅱ, treatment with a decoction of the Fuzhengzengxiao formula alone; Group Ⅲ, treatment with radiotherapy alone; and Group Ⅳ, treatment with radiotherapy plus a decoction of Fuzhengzengxiao formula. When the tumor body was 1 cm^3 in diameter, the tumor bearing mice in GroupsⅢ and Ⅳ were irradiated at a single dose of 10 Gy and the tumor inhibition rate was evaluated.The expression of S100A9 was determined using Western blotting and q-PCR(Real-time Quantitative PCR Detecting System). The sensitivity of cells containing RNAi S100A9 to radiotherapy was evaluated using the Click multiple target model,and the cell cycle was analyzed using flow cytometry.RESULTS: Relative to the control group,the expression of S100A9 in the tumors in each treatment group was decreased,especially in Group Ⅳ. The sensitizing enhancement ratio(SER) Dq was >1 after RNAi S100A9; it decreased the surviving fraction after a 2 Gy dose exposure,and also the D_0 and Dq of the tumor cells; in addition, the radiosensitivity of G_2/M cells was significantly increased.CONCLUSION: The decoction of the Fuzhengzengxiao formula downregulated the expression of S100A9 in lung adenocarcinoma cells.

Effect and mechanism of CyPA in radiosensitization of lung adenocarcinoma using CRISPR/Cas9 technology

Objective:To explore the role of cyclophilin A(CyPA)in sensitization of lung adenocarcinoma to radiotherapy using CRISPR/Cas9 technology.Methods:A CyPA knockout human lung adenocarcinoma cell line H1975 was established by CRISPR/Cas9 technology.Groups included a control group(wildtype),CyPA knockout group,traditional Chinese medicine(TCM)extract with Fuzhengzengxiao decoction group,and TCM extract with Fuzhengzengxiao decoction t CyPA knockout group.Each group was exposed to radiation at doses of 0,2,4,6,and 8 Gy.After 24 h,MTT assays were used to determine the survival rate of lung cancer cells and calculate radiosensitivity.The qPCR was used to measure mRNA expression of DDIT3,CDKN1A,and CDC25A associated with DNA damage repair.Results:Without irradiation,Fuzhengzengxiao decoction reduced the survival rate of lung adenocarcinoma cells(P<.0001).After irradiation,TCM extract with Fuzhengzengxiao decoction,CyPA knockout,and TCM extract with Fuzhengzengxiao decoction t CyPA knockout groups had reduced survival rates(P<.0001)and radiosensitivity was increased significantly.Expression of DDIT3,CDKN1A,and CDC25A was upregulated after knockout of CyPA(P<.0001).Expression of DDIT3 and CDC25A was increased after irradiation in wildtype cells treated with TCM extract with Fuzhengzengxiao decoction(DDIT3,P<.0001;CDC25A,P紏.0059).The TCM extract with Fuzhengzengxiao decoction t CyPA knockout group also had increased expression of DDIT3 and CDC25A after irradiation(P<.0001).Conclusion:Fuzhengzengxiao decoction significantly decreases the survival rate of lung cancer cells,and its mechanism may be related to radiosensitization by decreasing expression of CyPA and inducing G1/S cell cycle arrest.

扶正增效方放射增敏与肿瘤细胞内O^6-甲基鸟嘌呤DNA甲基转移酶的关系

目的探讨扶正增效方放射增敏与肿瘤细胞内 O^6-甲基鸟嘌呤 DNA 甲基转移酶(O^6-MT)关系。方法建立裸鼠人肺腺癌 PAa 模型,分为6组,从接种日始,Ⅱ、Ⅳ、Ⅵ组裸鼠每天灌服0.6g 扶正增效方水煎剂,Ⅰ、Ⅲ组灌服等量无菌生理盐水,35天Ⅴ、Ⅵ组腹腔注射2mg STZ,36天予Ⅲ、Ⅳ、Ⅴ、Ⅵ组瘤体10Gy X 线照射,测瘤体直径、O^6-MT 含量、MGMT 染色及细胞病理学镜检。结果 6组中O^6-MT 含量工组最高,Ⅵ组最少,Ⅳ、Ⅴ、Ⅵ组与Ⅰ组相比差异显著,Ⅳ、Ⅵ组相比差异也显著;MGMT 染色强度变化与 O^6-MT 含量一致。瘤体生长Ⅰ、Ⅱ组瘤体放疗第4天起明显增大,Ⅲ组第6天明显增大,与Ⅰ、Ⅱ组相比增长减慢;Ⅳ、Ⅴ、Ⅵ组瘤体在第4天明显减少,以第Ⅵ组最明显;病理学镜检Ⅲ组见小的散在坏死灶,Ⅳ、Ⅴ组坏死区域较Ⅲ组增加,Ⅳ组与Ⅴ组相似;Ⅵ组坏死区域明显增多增大。通过对 O^6-MT 活性、MGMT 染色强度与病理学镜检、瘤体大小对比,细胞内 O^6-MT 含量与放射敏感性密切相关,O^6-MT 含量低组别放疗效果好。结论 O^6-MT含量决定其放射敏感性,据 O^6-MT 活性高低预测其放射敏感性;扶正增效方放射增敏一个重要途径是抑制 DNA 甲基转移酶活性而实现的。