Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 703-712.DOI: 10.19852/j.cnki.jtcm.20240617.002
• Original articles • Previous Articles Next Articles
GUO Yuxi1, LI Ze1, CHENG Nan1, JIA Xuemei1, WANG Jie1, MA Hongyu2, ZHAO Runyuan3, LI Bolin1, XUE Yucong4, CAI Yanru1(), YANG Qian1(
)
Received:
2023-03-22
Accepted:
2023-07-24
Online:
2024-08-15
Published:
2024-06-17
Contact:
YANG Qian, Department of spleen and stomach diseases, First Affiliated Hospital of Hebei University of Traditional Chinese Medicine, Shijiazhuang 050000, China. Supported by:
GUO Yuxi, LI Ze, CHENG Nan, JIA Xuemei, WANG Jie, MA Hongyu, ZHAO Runyuan, LI Bolin, XUE Yucong, CAI Yanru, YANG Qian. High-throughput sequencing analysis of differential microRNA expression in the process of blocking the progression of chronic atrophic gastritis to gastric cancer by Xianglian Huazhuo formula (香连化浊方)[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 703-712.
Figure 1 Histologic changes in the gastric mucosal tissue (magnification × 200, scale bar 50 μm) A: normal group; B: model group; C: XLHZ group. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. HE: hematoxylin and eosin; CAG: chronic atrophic gastritis; XLHZ: Xianglian Huazhuo formula.
Group | n | Normal | Model | XLHZ |
---|---|---|---|---|
miR-20a-3p | 3 | 1.08±0.49 | 6.65±1.38a | 3.31±0.76b |
miR-320-3p | 3 | 1.04±0.31 | 0.15±0.06a | 0.44±0.15b |
miR-34b-5p | 3 | 1.05±0.37 | 0.11±0.07a | 0.58±0.13c |
miR-483-3p | 3 | 1.01±0.15 | 0.29±0.16a | 0.37±0.12 |
miR-883-3p | 3 | 1.03±0.28 | 0.21±0.05a | 0.52±0.17b |
Akap12 | 3 | 1.03±0.25 | 0.19±0.13a | 0.76±0.25b |
DLL1 | 3 | 1.00±0.11 | 0.32±0.07a | 0.58±0.06b |
NR4A2 | 3 | 1.06±0.40 | 3.28±0.50a | 2.25±0.49b |
TFRC | 3 | 1.01±0.18 | 2.13±0.29a | 1.64±0.22b |
Table 1 qRT-PCR validation of miRNAs expression
Group | n | Normal | Model | XLHZ |
---|---|---|---|---|
miR-20a-3p | 3 | 1.08±0.49 | 6.65±1.38a | 3.31±0.76b |
miR-320-3p | 3 | 1.04±0.31 | 0.15±0.06a | 0.44±0.15b |
miR-34b-5p | 3 | 1.05±0.37 | 0.11±0.07a | 0.58±0.13c |
miR-483-3p | 3 | 1.01±0.15 | 0.29±0.16a | 0.37±0.12 |
miR-883-3p | 3 | 1.03±0.28 | 0.21±0.05a | 0.52±0.17b |
Akap12 | 3 | 1.03±0.25 | 0.19±0.13a | 0.76±0.25b |
DLL1 | 3 | 1.00±0.11 | 0.32±0.07a | 0.58±0.06b |
NR4A2 | 3 | 1.06±0.40 | 3.28±0.50a | 2.25±0.49b |
TFRC | 3 | 1.01±0.18 | 2.13±0.29a | 1.64±0.22b |
Figure 2 Effect of XLHZ on proliferation and migration of MNNG-induced GES-1 cells A: EdU assay was used to detect cell proliferation (magnification × 200). The proliferating cells were fluorescently stained with EdU (Green). The nuclei were stained with DAPI (Blue). the higher the cell proliferation rate, the brighter the green in the plot. A1: EdU of the control group; A2: DAPI of the control group; A3: merge of the control group; A4: EdU of the model group; A5: DAPI of the model group; A6: merge of the model group; A7: EdU of the XLHZ group; A8: DAPI of the XLHZ group; A9: merge of the XLHZ group. B: Quantitative analysis of proliferating cells. C: Wound healing assay was used to detect cell migration in the normal, model and XLHZ groups (magnification × 200). C1-C5: The migration of cells in the normal group was observed at 0, 12, 24, 48, 72 h. C6-C10: The migration of cells in the model group was observed at 0, 12, 24, 48, 72 h. C11-C15: The migration of cells in the XLHZ group was observed at 0, 12, 24, 48, 72 h. D: quantitative analysis of cells migration rate at 0, 12, 24, 48, 72 h. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells; EdU: 5-Ethynyl-2’- deoxyuridine; DAPI: 4’,6-Diamidino-2’-phenylindole. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.05, bP < 0.01; Compared with the model group, cP < 0.05; dP < 0.01.
Figure 3 Effect of XLHZ on apoptosis and cell cycle of MNNG-induced GES-1 cells A: flow cytometry assay was used to detect apoptosis in the normal, model and XLHZ groups. A1: normal group; A2: model group; A3: XLHZ group; A4: apoptosis rate of quantitative analysis. B: flow cytometry assay was used to detect cell cycle in the normal, model and XLHZ groups. B1: normal group; B2: model group; B3: XLHA group; B4: quantitative analysis of cell cycle distribution. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.01; Compared with the model group, bP < 0.01.
Figure 4 Effects of XLHZ on EMT and proliferation of MNNG-induced GES-1 cells A: representative images of of E-cadherin, TGF-β1, vimentin and β-catenin protein expression that were exhibited from MNNG-induced GES-1 cells in the normal, model and XLHZ groups. B: quantitative analysis of E-cadherin protein; C: quantitative analysis of TGF-β1 protein; D: quantitative analysis of vimentin protein; E: quantitative analysis of β-catenin protein. normal group: drink and eat freely + the same volume of normal saline was given by gavage; model group: MNNG, 180 μg/mL + 2% sodium salicylate, 1 mL/100 g per day + hunger and satiety disorders; XLHZ: model group + XLHZ, 36 g·kg-1·d-1. XLHZ: Xianglian Huazhuo formula; EMT: epithelial mesenchymal transition; MNNG: N-Methyl-N'-Nitro-N-Nitrosoguanidine; GES-1: human gastric mucosal epithelial cells. Statistical analyses were measured using one-way analysis of variance followed by post hoc Bonferroni correction for multiple comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the normal group, aP < 0.01; Compared with the model group, bP < 0.01; cP < 0.05.
1. | Piscione M, Mazzone M, Di Marcantonio MC, et al. Eradication of Helicobacter pylori and gastric cancer: a controversial relationship. Front Microbiol 2021; 12: 630852. |
2. |
Ohata H, Kitauchi S, Yoshimura N, et al. Progression of chronic atrophic gastritis associated with Helicobacter pylori infection increases risk of gastric cancer. Int J Cancer 2004; 109: 138-43.
DOI PMID |
3. | Correa P. Human gastric carcinogenesis: a multistep and multifactorial process- -First American Cancer Society Award Lecture on Cancer Epidemiology and Prevention. Cancer Res 1992; 52: 6735-40. |
4. | Compare D, Rocco A, Nardone G. Risk factors In gastric cancer. Eur Rev Med Pharmacol Sci 2010; 14: 302-8. |
5. |
Taniguchi K, Karin M. NF-κB, inflammation, immunity and cancer: coming of age. Nat Rev Immunol 2018; 18: 309-24.
DOI PMID |
6. | Kaltschmidt B, Greiner JFW, Kadhim HM, et al. Subunit-specific role of NF-κB in cancer. Biomedicines 2018; 6: 44. |
7. |
Mayer RJ, Venook AP, Schilsky RL. Progress against GI cancer during the american society of clinical oncology's first 50 years. J Clin Oncol 2014; 32: 1521-30.
DOI PMID |
8. |
Rodriguez-Castro KI, Franceschi M, Noto A, et al. Clinical manifestations of chronic atrophic gastritis. Acta Biomed 2018; 89: 88-92.
DOI PMID |
9. |
Sharma PK, Suri TM, Venigalla PM, et al. Atrophic gastritis with high prevalence of Helicobacter pylori is a predominant feature in patients with dyspepsia in a high altitude area. Trop Gastroenterol 2014; 35: 246-51.
PMID |
10. |
Gu Z, Jia Q, Cong J, et al. Efficacy and safety of Elian Granules in treating chronic atrophic gastritis: study protocol for a randomized, double-blind, placebo-controlled, multicenter clinical trial. Trials 2022; 23: 437.
DOI PMID |
11. | Ou J, Wang L. Efficacy of self-made Hewei decoction for chronic atrophic gastritis and its effect on gastrin and pepsinogen expression levels. Contrast Media Mol Imaging 2022; 2022: 1092695. |
12. | Yang T, Wang R, Zhang J, et al. Mechanism of berberine in treating Helicobacter pylori induced chronic atrophic gastritis through IRF8-IFN-γ signaling axis suppressing. Life Sci 2020; 248: 117456. |
13. | Tong Y, Wang R, Liu X, et al. Zuojin pill ameliorates chronic atrophic gastritis induced by MNNG through TGF-β1/PI3K/Akt axis. J Ethnopharmacol 2021; 271: 113893. |
14. | Wang J, Gao YX, Ma HY, et al. Mechanism of Xianglian Huazhuo prescription against chronic atrophic gastritis based on network pharmacology and experimental verification. Zhong Guo Shi Yan Fang Ji Xue Za Zhi 2022; 28: 161-8. |
15. | Gao YX, Wang J, Ma HY, et al. Effects of Xianglian Huazhuo recipe on apoptosis signal pathway related factors in rats with chronic atrophic gastritis. Zhong Hua Zhong Yi Yao Za Zhi 2023; 38: 1050-7. |
16. | Wang J, Ma HY, Gao YX, et al. Protective effect of Huazhuo Jiedu recipe on chronicatrophic gastritis in rats. Zhong Guo Zhong Xi Yi Jie He Za Zhi 2022; 42: 348-54. |
17. |
Hu ML, Xiong SW, Zhu SX, et al. MicroRNAs in gastric cancer: from bench to bedside. Neoplasma 2019; 66: 176-86.
DOI PMID |
18. | Cheng J, Yang A, Cheng S, et al. Circulating miR-19a-3p and miR-483-5p as novel diagnostic biomarkers for the early diagnosis of gastric cancer. Med Sci Monit 2020; 26: e923444. |
19. |
Feng X, Zhu M, Liao B, et al. Upregulation of miR-552 predicts unfavorable prognosis of gastric cancer and promotes the proliferation, migration, and invasion of gastric cancer cells. Oncol Res Treat 2020; 43: 103-11.
DOI PMID |
20. | Xu J, Zhang Z, Chen Q, et al. miR-146b regulates cell proliferation and apoptosis in gastric cancer by targeting PTP1B. Dig Dis Sci 2020; 65: 457-63. |
21. |
Nardone G, Rocco A, Compare D, et al. Is screening for and surveillance of atrophic gastritis advisable? Dig Dis 2007; 25: 214-7.
PMID |
22. | Zhou Y, Xu Q, Shang J. et al, Crocin inhibits the migration, invasion,and epithelial-mesenchymal transition of gastric cancer cells via miR-320/KLF5/HIF-lα signaling. J Cell Physiol 2019; 234: 17876-85. |
23. | Song M, Wang X, Luo Y, et al. Cantharidin suppresses gastric cancer cell migration/invasion by inhibiting the PI3K/Akt signaling pathway via CCAT1, Chem Biol Interact 2020; 317: 108939. |
24. | Liu ZM, Yang XL, Jiang F, et al. Matrine involves in theprogression of gastric cancer through inhibiting miR-93-Sp and upregulating the expression of target gene AHNAK. J Cell Biochem 2020; 121: 2467-77. |
25. |
Tsukamoto H, Mizoshita T, Katano T, et al. Preventive effect of rebamipide on N-methyl-N'-nitro-N-nitrosoguanidine-induced gastric carcinogenesis in rats. Exp Toxicol Pathol 2015; 67: 271-7.
DOI PMID |
26. | Si J, Zhou W, Wu J, et al. Establishment of an animal model of chronic atrophic gastritis and a study on the factors inducing atrophy. Chin Med J (Engl) 2001; 114: 1323-5. |
27. | Adams BD, Kasinski AL, Slack FJ. Aberrant regulation and function of microRNAs in cancer. Curr Biol 2014; 24: R762-76. |
28. |
Seven M, Karatas OF, Duz MB, et al. The role of miRNAs in cancer: from pathogenesis to therapeutic implications. Future Oncol 2014; 10: 1027-48.
DOI PMID |
29. | Seeneevassen L, Bessède E, Mégraud F, et al. Gastric cancer: advances in carcinogenesis research and new therapeutic strategies. Int J Mol Sci 2021; 22: 3418. |
30. | Usman S, Waseem NH, Nguyen TKN, et al. Vimentin is at the heart of epithelial mesenchymal transition (EMT) mediated metastasis. Cancers (Basel) 2021; 13: 4985. |
31. | Danielsson F, Peterson MK, Caldeira Araújo H, et al. Vimentin diversity in health and disease. Cells 2018; 7: 147. |
32. | Liu CY, Lin HH, Tang MJ, et al. Vimentin contributes to epithelial-mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget 2015; 6: 15966-83. |
33. |
Wenzel ES, Singh ATK. Cell-cycle checkpoints and aneuploidy on the path to cancer. In Vivo 2018; 32: 1-5.
PMID |
34. | Khan M, Rasul A, Yi F, et al. Jaceosidin induces p53-dependent G2/M phase arrest in U87 glioblastoma cells. Asian Pac J Cancer Prev 2011; 12: 3235-8. |
35. |
Icard P, Fournel L, Wu Z, et al. Interconnection between metabolism and cell cycle in cancer. Trends Biochem Sci 2019; 44: 490-501.
DOI PMID |
36. | Wang Y, Zhou X, Shan B, et al. Downregulation of microRNA-33a promotes cyclin-dependent kinase 6, cyclin D1 and PIM1 expression and gastric cancer cell proliferation. Mol Med Rep 2015; 12: 6491-500. |
37. | Xu X, Lai Y, Hua ZC. Apoptosis and apoptotic body: disease message and therapeutic target potentials. Biosci Rep 2019; 39: BSR20180992. |
38. |
Wei S, Peng L, Yang J, et al. Exosomal transfer of miR-15b-3p enhances tumorigenesis and malignant transformation through the DYNLT1/caspase-3/caspase-9 signaling pathway in gastric cancer. J Exp Clin Cancer Res 2020; 39: 32.
DOI PMID |
[1] | SHEN Jie, YIN Yaoli, LI Hongxiao, LU Ge, ZHU Yaoyao, QIN Yantong, JIN Xun, CHENG Jie, SHEN Meihong. Effect of moxibustion on expression profile of miRNAs in Tripterygium glycoside-induced decreased ovarian reserve [J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 745-752. |
[2] | DING Luobin, WANG Huajun, LI Yao, LI Jia, LI Ling, GAO Yangping, GUAN Jian, GENG Weiqiang. Electroacupuncture stimulating Neixiyan (EX-LE5) and Dubi (ST35) alleviates osteoarthritis in rats induced by anterior cruciate ligament transaction via affecting DNA methylation regulated transcription of miR-146a and miR-140-5p [J]. Journal of Traditional Chinese Medicine, 2023, 43(5): 983-990. |
[3] | HOU Chao, ZHANG Yusen, YANG Die, LI Yifei, ZHANG Xiaochun, LIU Yanqing. Effects of Traditional Chinese Medicine on the survival of patients with stage I gastric cancer and high-risk factors: a real-world retrospective study [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 568-573. |
[4] | DING Yajie, LIU Feng, LI Zhaoyan, Xu Yan, CAO Nida, ZHANG Guangao, WANG Rui, ZHAO Aiguang. Efficacy of luteolin on the human gastric cancer cell line MKN45 and underlying mechanism [J]. Journal of Traditional Chinese Medicine, 2023, 43(1): 34-41. |
[5] | XU Qing, WANG Yang, LI Zhongyu, YAN Jiaxing, ZHAO Yingpan, WANG Ping, WEN Yandong. Therapeutic mechanisms of integrated traditional Chinese and conventional medicine underlying its treatment of precancerous lesions of gastric cancer [J]. Journal of Traditional Chinese Medicine, 2022, 42(6): 1023-1028. |
[6] | DING Jiamin, XING Yifeng, CHEN Zuoliang, CHEN Wanlu, MA Zhongxiong, XIE Yunde, ZHOU Lin. Qilan preparation (芪蓝颗粒) inhibits proliferation and induces apoptosis by down-regulating microRNA-21 in human Tca8113 tongue squamous cell carcinoma cells [J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 693-700. |
[7] | PAN Lijia, MA Shuya, WEN Jing, ZHANG Xiaoqi, XING Haijiao, JIA Chunsheng. Direct contact moxibustion promotes apoptosis of gastric cancer cells in rats by regulating intestinal flora [J]. Journal of Traditional Chinese Medicine, 2021, 41(6): 943-952. |
[8] | Wu Jiarui, Zhang Dan, Ni Mengwei, Xue Jiaping, Wang Kaihuan, Duan Xiaojiao, Liu Shuyu. Effectiveness of Huachansu injection combined with chemotherapy for treatment of gastric cancer in China: a systematic review and Meta-analysis [J]. Journal of Traditional Chinese Medicine, 2020, 40(5): 749-757. |
[9] | Gao Lujiao, Niu Xin, Niu Tingli, Wang Xuan, Lu Xiaoyan, Feng Qianjin, Yang Xuezhi. Curative efficacy of extract from Ganjiangdazao recipe on functional dyspepsia in rats [J]. Journal of Traditional Chinese Medicine, 2020, 40(4): 654-663. |
[10] | Deng Xin, Liang Xingqiu, Zhou Xiaoxiao, Jiang Manjun, Liang Mingkun, Wang Xinyuan, Zhao Xiaofang, Fu Lei, Liang Jian. Protective effect and mechanisms of Weining granule on N-methyl-N'-nitro-N-nitrosoguanidine-induced gastric cancer in rats [J]. Journal of Traditional Chinese Medicine, 2019, 39(03): 393-401. |
[11] | Cao Yunxiang, Guo Yunke, Zong Ruikai, Huang Chuanbing, Wang Yue, Liu Jian. Drug-containing serum of Xinfeng capsules protect against H9C2 from death by enhancing miRNA-21 and inhibiting toll-like receptor 4/phosphorylated p-38(p-p38)/p-p65 signaling pathway and proinflammatory cytokines expression [J]. Journal of Traditional Chinese Medicine, 2018, 38(03): 359-365. |
[12] | Wang Hongwei, Li Hailong, Chen Fengqin, Luo Jun, Gu Jing, Wang Huping, Wu Hongyan, Xu Yan. Baicalin extracted from Huangqin(Radix Scutellariae Baicalensis) induces apoptosis in gastric cancer cells by regulating B cell lymphoma(Bcl-2)/Bcl-2-associated X protein and activating caspase-3 and caspase-9 [J]. Journal of Traditional Chinese Medicine, 2017, 37(02): 229-235. |
[13] | Wang Jiancheng, Ge Long, Zhao Ye, Li Jinlong, Zhang Pan, Mao Lei, Yang Kehu. Effect of Aidi injection plus chemotherapy on gastric carcinoma:a Meta-analysis of randomized controlled trials [J]. Journal of Traditional Chinese Medicine, 2015, 35(04): 361-374. |
[14] | Junfeng Zhang, Zhen Zhan, Juan Wu, Chunbing Zhang, Yaping Yang, Shujuan Tong, Ruiping Wang, Xuewen Yang, Wei Dong. Association among lifestyle, clinical examination, polymorphisms in CDH1 gene and Traditional Chinese Medicine syndrome differentiation of gastric cancer [J]. Journal of Traditional Chinese Medicine, 2013, 33(05): 572-579. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||
Sponsored by China Association of Chinese Medicine
& China Academy of Chinese Medical Sciences
16 Nanxiaojie, Dongzhimen Nei, Beijing, China. 100700 Email: jtcmen@126.com
Copyright 2020 Journal of Traditional Chinese Medicine. All rights reserved.