Tenglong Buzhong granules (藤龙补中颗粒) inhibits the growth of SW620 human colon cancer in vivo

doi:10.19852/j.cnki.jtcm.2022.05.004

Abstract

Abstract:

OBJECTIVE: To observe the anticancer effects of the granular preparation of Tenglong Buzhong decoction (藤龙补中汤,TBD), i.e Tenglong Buzhong granules (藤龙补中颗粒, TBG), in human SW620 colon cancer.

METHODS: BALB/c nude mice were subcutaneously transplanted with SW620 cells, and treated with TBG (2.56 g/kg, once per day) and/or 5-Fu (104 mg/kg, once per week) for 21 d. Apoptosis, Caspase activities and cellular senescence were measured by commercial kits. The protein expression and phosphorylation were detected by Western blot or immunohistochemistry.

RESULTS: TBG and 5-Fu inhibited tumor growth. The tumor inhibition rate of the TBG, 5-Fu, and TBG+5-Fu groups was 42.25%, 51.58%, and 76.08%, respectively. Combination of TBG and 5-Fu showed synergetic anti-cancer effects. TBG and 5-Fu induced apoptosis, activated caspase-3, -8, and -9, increased SMAC expression, inhibited XIAP expression. TBG induced cellular senescence, upregulated cyclin-dependent kinase inhibitor 1a (CDKN1a) and cyclin-dependent kinase inhibitor 2a (CDKN2a) expression, and inhibited phosphorylation of retinoblastoma-associated protein (RB) and expression of cyclin E1 (CCNE1) and cyclin-dependent kinases (CDK) 2. TBG also inhibited angiogenesis accompanied by downregulation of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α).

CONCLUSIONS: TBG inhibits SW620 colon cancer growth, induces apoptosis via SMAC-XIAP-Caspases signaling, induces cellular senescence through CDKN1a/CDKN2a-RB-E2F signaling, inhibits angiogenesis by down-regulation of HIF-1α and VEGF, and enhances the effects of 5-Fu.

Key words: colonic neoplasms, apoptosis, cellular senescence, signal transduction, angiogenesis, Tenglong Buzhong granules

LI Miao, ZHENG Jialu, WANG Shuangshuang, CHEN Lei, PENG Xiao, CHEN Jinfang, AN Hongmei, HU Bing. Tenglong Buzhong granules (藤龙补中颗粒) inhibits the growth of SW620 human colon cancer in vivo[J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 701-706.

Figures/Tables 4

Figure 1 Effects of TBG on tumor growth A: tumor volume; B: tumor weight; C: mice body weight; D: Ki-67 expression was detected by immunohistochemistry (×200); E: quantification of Ki-67 expression; F: hematoxylin-eosin staining of tumor tissues (×200). D1 and F1, control group; D2 and F2, 5-Fu group; D3 and F3, TBG group; D4 and F4, TBG+5-Fu group. aP < 0.01 vs control group; bP < 0.05, cP < 0.01 vs TBG or 5-Fu group. TBG, Tenglong Buzhong granules.

Figure 2 TBG induced apoptosis A: apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) (×200); B: the apoptotic cells were counted by Image Pro 6.0 software; C: caspases activities; D: cleavage of PARP was detected by Western blot; E: protein expression was detected by Western blot; F: quantification of protein expression. A1, control group; A2, 5-Fu group; A3, TBG group; A4, TBG+5-Fu group. aP < 0.01 vs control group; bP < 0.01, cP < 0.05 vs TBG or 5-Fu group. TBG: Tenglong Buzhong granules; c-PARP: cleaved poly ADP-ribose polymerase; GAPDH: glyceraldehyde-3-phosphatedehydrogenase; SMAC: second mitochondria-derived activator of caspases; XIAP: X-linked inhibitor of apoptosis protein.

Figure 3 TBG induced cellular senescence A: senescence-associated β-galactosidase (SA-β-Gal) staining (×200); A1: control group; A2: 5-Fu group; A3: TBG group; A4: TBG+5-Fu group; B: quantification of positive SA-β-Gal staining. C-E, protein expression and phosphorylation was detected by Western blot and quantified. aP > 0.05, bP < 0.01, vs control group; cP < 0.01 vs 5-Fu group; dP > 0.05 vs TBG group. TBG: Tenglong Buzhong granules; CDKN1a: cyclin-dependent kinase inhibitor 1a; CDKN2a: cyclin-dependent kinase inhibitor 2a; RB: retinoblastoma-associated protein; pRB: phosphorylated RB; GAPDH: glyceraldehyde-3-phosphatedehydrogenase; E2F1: E2F transcription factor 1; CDK2: cyclin-dependent kinase 2; CCNE1: cyclin E1.

Figure 4 TBG inhibited angiogenesis A: angiogenesis was identified by immunohistochemistry with CD31 antibody (×200); A1: control group; A2: 5-Fu group; A3: TBG group; A4: TBG+5-Fu group; B: quantification of angiogenesis. C, D: protein expression was detected by Western blot and quantified. TBG: Tenglong Buzhong granules; VEGF: Vascular endothelial growth factor; HIF-1α: Hypoxia-inducible factor-1α; GAPDH: Glyceraldehyde-3-phosphatedehydrogenase. aP > 0.05, bP < 0.01, vs control group; cP < 0.01 vs 5-Fu group; dP > 0.05 vs TBG group.

References 35.

1.	Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71: 209-49. DOI URL
2.	Cohen R, Pudlarz T, Delattre JF, et al. Molecular targets for the treatment of metastatic colorectal cancer. Cancers (Basel) 2020; 12: 2350. DOI URL
3.	Loree JM, Kopetz S. Recent developments in the treatment of metastatic colorectal cancer. Ther Adv Med Oncol 2017; 9: 551-64. DOI URL
4.	Kong MY, Li LY, Lou YM, et al. Chinese herbal medicines for prevention and treatment of colorectal cancer: from molecular mechanisms to potential clinical applications. J Integr Med 2020; 18: 369-84. DOI URL
5.	Deng S, Hu B, An HM. Traditional Chinese medicinal syndromes and treatment in colorectal cancer. J Cancer Ther 2012; 3: 888-97. DOI URL
6.	Wang T, Nan H, Zhang C, et al. Aidi injection combined with FOLFOX 4 chemotherapy regimen in the treatment of advanced colorectal carcinoma. J Cancer Res Ther 2014; Suppl 1: 52-5.
7.	Yang M, Zhu SJ, Shen C, et al. Clinical application of Chinese herbal injection for cancer care: evidence-mapping of the systematic reviews, Meta-analyses, and randomized controlled trials. Front Pharmacol 2021; 12:666368. DOI URL
8.	Gan C, Jin Z, Wei X, et al. Actinidia chinensis Planch. root extract inhibits the proliferation, migration and invasion of breast cancer cells via the AKT/GSK-3β signaling pathway. Folia Histochem Cytobiol 2021; 59: 226-35. DOI URL
9.	Gao Z, Deng G, Li Y, et al. Actinidia chinensis Planch prevents proliferation and migration of gastric cancer associated with apoptosis, ferroptosis activation and mesenchymal phenotype suppression. Biomed Pharmacother 2020; 126: 110092. DOI URL
10.	Ling B, Xiao S, Yang J, et al. Probing the antitumor mechanism of Solanum nigrum L. aqueous extract against human breast cancer MCF7 cells. Bioengineering (Basel) 2019; 6: 112.
11.	Yan X, Li M, Chen L, et al. α‑Solanine inhibits growth and metastatic potential of human colorectal cancer cells. Oncol Rep 2020; 43: 1387-96.
12.	Yue GG, Chan YY, Liu W, et al. Effectiveness of Scutellaria barbata water extract on inhibiting colon tumor growth and meta-stasis in tumor-bearing mice. Phytother Res 2021; 35: 361-73. DOI URL
13.	Jin Y, Chen W, Yang H, et al. Scutellaria barbata D. Don inhibits migration and invasion of colorectal cancer cells via suppression of PI3K/AKT and TGF-β/Smad signaling pathways. Exp Ther Med 2017; 14: 5527-34.
14.	Mao W, Fan Y, Cheng C, et al. Efficacy and safety of Kanglaite injection combined with chemotherapy for colorectal cancer: a protocol for systematic review and Meta-analysis. Medicine (Baltimore) 2020; 99: e22357. DOI URL
15.	Hu B, Li G, An H, et al. Clinical trial of Tenglong Buzhong decoction plus chemotherapy for metastatic colorectal cancer. Zhong Hua Zhong Yi Yao Xue Kan 2015; 33: 37-9.
16.	Hu B, Li G, An H, et al. Effects of Tenglong Buzhong decoction on Th1 immune response in patients with advanced colorectal cancer. Zhong Guo Zhong Xi Yi Jie He Xiao Hua Za Zhi 2014; 22: 434-7.
17.	Hu B, An HM, Shen KP, et al. Effects of Tenglong Buzhong decoction on proliferation and apoptosis of human colon carcinoma cell line LS174T. Zhong Xi Yi Jie He Xue Bao 2010; 8: 575-80. DOI URL
18.	Hu B, An HM, Shen KP, et al. Senescence-inducing effects of Chinese herbal medicine Tenglong Buzhong decoction on human colon carcinoma LS-174-T cells and the mechanism. Zhong Xi Yi Jie He Xue Bao 2010; 8: 1048-52. DOI URL
19.	Xu C, Wang X, Liang H, et al. Optimization of water extraction process for Tenglong Buzhong recipe by orthogonal test. Zhong Guo Yao Ye 2019; 28: 15-8.
20.	Foucquier J, Guedj M. Analysis of drug combinations: current methodological landscape. Pharmacol Res Perspect 2015; 3: e00149. DOI URL
21.	Koff JL, Ramachandiran S, Bernal-Mizrachi L. A time to kill: targeting apoptosis in cancer. Int J Mol Sci 2015; 16: 2942-55. DOI URL
22.	Cossu F, Milani M, Mastrangelo E, et al. Targeting the BIR domains of inhibitor of apoptosis (IAP) proteins in cancer treatment. Comput Struct Biotechnol J 2019; 17: 142-50. DOI URL
23.	Cairney CJ, Bilsland AE, Evans TR, et al. Cancer cell senescence: a new frontier in drug development. Drug Discov Today 2012; 17: 269-76. DOI PMID
24.	Chiantore MV, Vannucchi S, Mangino G, et al. Senescence and cell death pathways and their role in cancer therapeutic outcome. Curr Med Chem 2009; 16: 287-300. PMID
25.	Dimri GP, Lee X, Basile G, et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 1995; 92: 9363-67. DOI URL
26.	Yang MY, Hung CH, Chang CH, et al. Solanum nigrum suppress angiogenesis-mediated tumor growth through inhibition of the AKT/mTOR pathway. Am J Chin Med 2016; 44: 1273-88. DOI URL
27.	Wei L, Lin J, Xu W, et al. Scutellaria barbata D. Don inhibits tu-mor angiogenesis via suppression of Hedgehog pathway in a mo-use model of colorectal cancer. Int J Mol Sci 2012; 13: 9419-30. DOI URL
28.	Tang RZ, Li ZZ, Hu D, et al. Sanjie Yiliu formula inhibits colorectal cancer growth by suppression of proliferation and induction of apoptosis. ACS Omega 2021; 6: 7761-70. DOI URL
29.	Dong B, Yang Z, Ju Q, et al. Anticancer effects of Fufang Yiliu Yin formula on colorectal cancer through modulation of the PI3K/Akt pathway and BCL-2 family Proteins. Front Cell Dev Biol 2020; 8: 704. DOI URL
30.	Yang H, Liu JX, Shang HX, et al. Qingjie Fuzheng granules inhibit colorectal cancer cell growth by the PI3K/AKT and ERK pathways. World J Gastrointest Oncol 2019; 11:377-92. DOI URL
31.	Tu H, Costa M. XIAP's profile in human cancer. Biomolecules 2020; 10: E1493.
32.	Lopez A, Harada K, Vasilakopoulou M, et al. Targeting angiogenesis in colorectal carcinoma. Drugs 2019; 79: 63-74. DOI URL
33.	Ioannou M, Paraskeva E, Baxevanidou K, et al. HIF-1α in colorectal carcinoma: review of the literature. J BUON 2015; 20: 680-9. PMID
34.	Tirpe AA, Gulei D, Ciortea SM, et al. Hypoxia: overview on hypoxia-mediated mechanisms with a focus on the role of HIF genes. Int J Mol Sci 2019; 20: 6140. DOI URL
35.	Lee SH, Jeong D, Han YS, et al. Pivotal role of vascular endothelial growth factor pathway in tumor angiogenesis. Ann Surg Treat Res 2015; 89: 1-8 DOI URL

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