Journal of Peking University (Health Sciences) ›› 2020, Vol. 52 ›› Issue (2): 247-253. doi: 10.19723/j.issn.1671-167X.2020.02.009

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Effect of Fei-Liu-Ping ointment combined with cyclophosphamide on lung cancer cell proliferation and acidic microenvironment

Liang GENG1,Jing LV2,(),Jing FAN3   

  1. 1. Department of Chinese and Western Medicine, Cancer Hospital Affiliated to Zhengzhou University, Zhengzhou 450008, China
    2. Traditional Chinese Medicine Teaching and Research Office, Zhengzhou Health Vocational College, Zhengzhou 450005, China
    3. College of Management, Henan University of Chinese Medicine, Zhengzhou 450008, China
  • Received:2019-10-25 Online:2020-04-18 Published:2020-04-18
  • Contact: Jing LV E-mail:1010489902@qq.com
  • Supported by:
    Supported by the National Natural Science Foundation of China and Science(81473638);Technology Project of Henan Province(172102310104)

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Abstract:

Objective: To observe the effects of Fei-Liu-Ping ointment and chemotherapy on mice with lung cancer, and to explore the inherent mechanism of action from the point of acidic microenvironment and apoptosis.Methods: First of all, the Lewis lung cancer transplanted mouse model was established. Therefore, they were treated by Fei-Liu-Ping ointment, cyclophosphamide, Fei-Liu-Ping ointment + cyclophosphamide and the saline as control. All the groups' tumor size, tumor growth rate and food consumption were recorded. The mice were sacrificed and the tumors were took out after 15 days' interventions. Then lactate relative concentrations were detected with lactate kits and the protein expressions of glucose transporter 4 (GLUT4), hexokinase 1 (HK1), glucose-regulated protein 78 (GRP78), carbo-nic anhydrase-Ⅸ (CA-Ⅸ) were detected through immunohistochemical staining. Flow cytometry was adopted to detect the percentage of apoptotic tumor cells and regulatory T cells (Treg), and the expression of hypoxia-inducible factor-1α (HIF-1α), Bcl-2, Bax, Caspase-3, interleukin-2 (IL-2) were tested through western blot.Results: The strongest inhibition effect and the lowest tumor growth rate was found in Fei-Liu-Ping ointment + cyclophosphamide group. There were significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group(P<0.05). And the highest food consumption was found in Fei-Liu-Ping ointment + cyclophosphamide group while there were no significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group (P>0.05). Further molecular biological detections found that the lowest lactate level and regulatory T cells ratio were found in Fei-Liu-Ping ointment + cyclophosphamide group and these expressions of GLUT4, HK1, GRP78, CA-Ⅸ were suppressed. There were significant differences between Fei-Liu-Ping ointment+cyclophosphamide group and saline group (P<0.05). In addition, the Fei-Liu-Ping ointment + cyclophosphamide group's cell apoptosis increased significantly compared with saline group and there were significant differences on expressions of HIF-1α, Bcl-2, Bax, Caspase-3, IL-2 for this group compared with saline group.Conclusion: Chemotherapy and Fei-Liu-Ping ointment had the synergistic effect on inhibiting tumor growth and improving the general conditions of tumor-bearing mice. The effect was partly owed to the improvement on tissue hypoxia, the inhibition of HIF-1α expression and the regulations on its downstream proteins, such as GLUT4, HK1, GRP78, and CA-Ⅸ. And then all these alterations led to the modulation tumor acidic microenvironment, the induced tumor cells apoptosis and suppression of T cells to regulatory T cells differentiation.

Key words: Lung neoplasms, Fei-Liu-Ping ointment, Chemotherapy, Tumor microenvironment, Apoptosis

CLC Number: 

  • R734.2

Figure 1

Mice's tumor size (A), tumor growth rate (B), food consumption (C) at different times after administration S, saline; F, Fei-Liu-Ping ointment; H, cyclophosphamide; H+F, cyclophosphamide + Fei-Liu-Ping ointment."

Figure 2

Lactate relative concentration in mice's tumor tissue * P<0.05, vs. S group."

Figure 3

CA-Ⅸ, GLUT4, GRP78 and HK1 protein expression in mice's tumor tissue (immunohistochemical staining ×40) S, saline; F, Fei-Liu-Ping ointment; H, cyclophosphamide; H+F, cyclophosphamide + Fei-Liu-Ping ointment. *P<0.05, vs. S group; #P<0.05, vs. H group."

Table 1

Apoptosis percent of tumour cells /%"

Group Early apoptosis Late apoptosis Total apoptosis
S 1.72±0.17 1.52±0.22 3.24±0.18
F 1.79±0.11 1.45±0.19 3.24±0.16
H 2.78±0.34 2.24±0.20 5.02±0.29
H+F 5.42±0.82 7.9±1.04 13.32±0.95*

Figure 4

Apoptosis percent of tumor cells"

Figure 5

HIF-1α, Bcl-2, Caspase-3, Bax and IL-2 protein expression in mice's tumor tissue A, results of Western blot; B-F, gray value. *P<0.05, vs. S group; #P<0.05, vs. H group."

Figure 6

Percent of Treg cells in mice's tumor*P<0.05, vs. S group."

[1] 陈璐, 高威 . 肿瘤酸性微环境的形成机制及其对肿瘤进展的影响[J]. 肿瘤, 2019,39(2):140-145.
[2] Li X, Yu X, Dai D , et al. The altered glucose metabolism in tumor and a tumor acidic microenvironment associated with extracellular matrix metalloproteinase inducer and monocarboxylate transporters[J]. Oncotarget, 2016,7(17):23141-23155.
[3] Wojtkowiak JW, Verduzco D, Schramm KJ , et al. Drug resistance and cellular adaptation to tumor acidic pH microenvironment[J]. Mol Pharm, 2011,8(6):2032-2038.
[4] 龚巍 . 调节性T细胞和胃癌的临床与实验研究[D]. 苏州: 苏州大学, 2017.
[5] Fukumura D, Jain RK . Tumor microvasculature and microenvironment: Targets for anti-angiogenesis and normalization[J]. Microvasc Res, 2007,74(2-3):72-84.
[6] Ganapathy V, Thangaraju M, Prasad PD . Nutrient transporters in cancer: Relevance to Warburg hypojournal and beyond[J]. Pharmacol Ther, 2009,121(1):29-40.
[7] Seagroves TN, Ryan HE, Lu H , et al. Transcription factor HIF-1 is a necessary mediator of the Pasteur effect in mammalian cells[J]. Mol Cell Biol, 2001,21(10):3436-3444.
[8] Semenza GL . Targeting HIF-1 for cancer therapy[J]. Nat Rev Cancer, 2003,3(10):721-732.
[9] Lee JW, Bae SH, Jeong JW , et al. Hypoxia-inducible factor (HIF-1) alpha: Its protein stability and biological functions[J]. Exp Mol Med, 2004,36(1):1-12.
[10] Winum JY, Rami M, Scozzafava A , et al. Carbonic anhydrase Ⅸ: A new druggable target for the design of antitumor agents[J]. Med Res Rev, 2008,28(3):445-463.
[11] van den Beucken T, Ramaekers CH, Rouschop K , et al. Deficient carbonic anhydrase 9 expression in UPR-impaired cells is associated with reduced survival in an acidic microenvironment[J]. Radiother Oncol, 2009,92(3):437-442.
[12] Verras M, Papandreou I, Lim AL , et al. Tumor hypoxia blocks Wnt processing and secretion through the induction of endoplasmic reticulum stress[J]. Mol Cell Biol, 2008,28(23):7212-7224.
[13] 张翔云, 周华妙, 郭勇 . 黄连解毒汤等对乳腺癌荷瘤小鼠肿瘤酸性微环境pH值的影响[J]. 黑龙江中医药, 2016,45(4):64-65.
[14] Flinck M, Kramer SH, Pedersen SF . Roles of pH in control of cell proliferation[J]. Acta Physiol (Oxf), 2018,223(3):e13068.
[15] Peppicelli S, Andreucci E, Ruzzolini J . The acidic microenvironment as a possible niche of dormant tumor cells[J]. Cell Mol Life Sci, 2017,74(15):2761-2771.
[16] Ryder C , McColl K,Zhong F, et al.Acidosis promotes Bcl-2 family-mediated evasion of apoptosis: Involvement of acid-sensing G protein-coupled receptor Gpr65 signaling to Mek/Erk[J]. J Biol Chem, 2012,287(33):27863-27867.
[17] Dhup S, Dadhich RK, Porporato PE , et al. Multiple biological activities of lactic acid in cancer: Influences on tumor growth, angiogenesis and metastasis[J]. Curr Pharm Des, 2012,18(10):1319-1330.
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