Procaine inhibits the proliferation, migration and invasion of gastric cancer cells through up-regulating lncRNA DGCR5

doi:10.3877/cma.j.issn.2095-1221.2024.03.004

Abstract

Abstract:

Objective

To explore procaine's role and potential mechanism in the migration, invasion and proliferation of gastric cancer cells.

Methods

Gastric cancer AGS cells were divided into low, medium, and high dose of procaine groups (cells were cultured in medium containing 3, 6, 12 mmol/L procaine for 24 h) , pcDNA-lncRNA DGCR5 group (AGS cells transfected with lncRNA DGCR5 overexpression vector were cultured in the conventional medium for 24 h) , pcDNA-NC group (AGS cells transfected with empty vector were cultured in the conventional medium for 24 h) , high-dose+si-lncRNA DGCR5 group (AGS cells transfected with lncRNA DGCR5 small interfering RNA were cultured in medium containing 12 mmol/L procaine for 24 h) , high-dose+si-NC group (AGS cells transfected with out-of-order nonsense negative sequence were cultured in medium containing 12 mmol/L procaine for 24 h) , and control group (AGS cells were cultured in conventional medium for 24 h) . And then, MTT, clone formation, scratch healing, Transwell and Western blot method were used to detect cell proliferation, migration and invasion, and the protein expression of MMP2 and MMP9 in cells, respectively; RT-qPCR was used to detected the expression of lncRNA DGCR5. Independent sample t-test was used for comparison between two groups, and univariate analysis of variance and LSD-t test were used for comparison between multiple groups.

Results

Compared with the control group, the A value, clone count, scratch healing rate, the number of migration and invasion of AGS cells, and the protein expression of MMP2 and MMP9 in the low, medium and high dose groups of procaine were decreased (P all < 0.05) , and the expression of lncRNA DGCR5 (1.69 ± 0.13, 2.18 ± 0.17, 2.49 ± 0.15 vs 1.00 ± 0.10) was increased (P < 0.05) . Compared with the pcDNA-NC group, the A value, clone count, scratch healing rate, the number of migration and invasion of AGS cells, and the protein expression of MMP2 and MMP9 in the pcDNA-lncRNA DGCR5 group were all decreased (P all < 0.05) . Compared with the high-dose+si-NC group, the A value, clone count, scratch healing rate, the number of migration and invasion of AGS cells, and the protein expression of MMP2 and MMP9 in the pcDNA-lncRNA DGCR5 group were all increased (P all < 0.05) .

Conclusion

Procaine may inhibit the migration, invasion proliferation and gastric cancer AGS cells by increasing the expression of lncRNA DGCR5.

Key words: Procaine, Gastric cancer, lncRNA DGCR5, Cell proliferation, Migration, Invasion

Jing Li, Xia Pan, Fang Zhou, Jing Wang, Jia Hong. Procaine inhibits the proliferation, migration and invasion of gastric cancer cells through up-regulating lncRNA DGCR5[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(03): 151-158.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2024.03.004

https://zhxbygxbzz.cma-cmc.com.cn/EN/Y2024/V14/I03/151

Figures/Tables 16

References 32

1	何家璇, 白洁, 李伟. 局部麻醉剂通过丝裂原活化蛋白激酶途径诱导人甲状腺癌细胞凋亡[J]. 临床和实验医学杂志, 2020, 19(3):263-267.
2	陈文华, 陈铭君, 张晓英, 等. 肺癌患者围术期医院感染危险因素及常用麻醉药物对预后的影响[J]. 中国医药, 2017, 12(9):1321-1325.
3	Bhol CS, Mishra SR, Patil S, et al. PAX9 reactivation by inhibiting DNA methyltransferase triggers antitumor effect in oral squamous cell carcinoma[J]. Biochim Biophys Acta Mol Basis Dis, 2022, 1868(9):166428. doi: 10.1016/j.bbadis.2022.166428.
4	Li C, Gao SH, Li XP, et al. Procaine inhibits the proliferation and migration of colon cancer cells through inactivation of the ERK/MAPK/FAK pathways by regulation of RhoA[J]. Oncol Res, 2018, 26(2):209-217.
5	曾凯辉, 吕慧莹, 罗金泰, 等. 普鲁卡因调控CXCR7并影响AKT和STAT3信号通路从而抑制膀胱癌细胞活力,迁移和侵袭[J]. 中国病理生理杂志, 2020, 36(4):612-618.
6	方洁, 朱建坡, 张虎, 等. 普鲁卡因通过调节SRY相关高迁移率族盒蛋白9抑制乳腺癌细胞增殖并诱导凋亡的机制研究[J]. 癌症进展, 2020, 18(15):1551-1555.
7	Li YC, Wang Y, Li DD, et al. Procaine is a specific DNA methylation inhibitor with anti-tumor effect for human gastric cancer[J]. J Cell Biochem, 2018, 119(2):2440-2449.
8	Xu Y, Zhang GH, Zou C, et al. Long noncoding RNA DGCR5 suppresses gastric cancer progression by acting as a competing endogenous RNA of PTEN and BTG1[J]. J Cell Physiol, 2019, 234(7):11999-12010.
9	韦玲, 杨凯, 刘焕, 等. 普鲁卡因调节lncRNA TTN-AS1表达对胃癌细胞恶性生物学行为的影响[J]. 河北医药, 2022, 44(10):1470-1474.
10	Herencia C, Diaz-Tocados JM, Jurado L, et al. Procaine inhibits osteo/odontogenesis through Wnt/β-Catenin inactivation[J]. PLoS One, 2016, 11(6):e0156788. doi: 10.1371/journal.pone.0156788.
11	Wang XL, Shi M, Xiang T, et al. Long noncoding RNA DGCR5 represses hepatocellular carcinoma progression by inactivating Wnt signaling pathway[J]. J Cell Biochem, 2019, 120(1):275-282.
12	朱晨宇, 高日文, 杜清, 等. 普鲁卡因对人肝癌HepG2细胞增殖影响的实验研究[J]. 现代肿瘤医学, 2012, 20(4):686-689.
13	马宝丰, 李铁成, 徐芳. 普鲁卡因通过miR-15a-5p/PI3K-AKT3途径调控乳腺癌细胞增殖、迁移和侵袭的机制[J]. 中国老年学杂志, 2020, 40(3):610-614.
14	方洁，朱建坡，张虎, 等. 普鲁卡因联合肌成束蛋白1对膀胱癌细胞的影响及机制研究[J]. 癌症进展, 2019, 17(13):1561-1564,1578.
15	Cai PY, Wu MX, Zhang B, et al. Long non-coding RNA SNHG12 regulates cell proliferation, invasion and migration in endometrial cancer by targeting miR-4429[J]. Mol Med Rep, 2020, 22(4):2842-2850.
16	Huang F, Jiang JJ, Yao YL, et al. Circular RNA Hsa_circRNA_101996 promotes the development of gastric cancer via upregulating matrix metalloproteinases-2/matrix metalloproteinases-9 through microRNA-143/Ten-eleven translocation-2 pathway[J]. J Cancer, 2021, 12(22):6665-6675.
17	Wang KH, Yu XY, Tao BL, et al. Downregulation of lncRNA HCP5 has inhibitory effects on gastric cancer cells by regulating DDX21 expression[J]. Cytotechnology, 2021, 73(1):1-11.
18	Pan C, Feng YH, Zhou J, et al. lncRNA HOTAIR knockdown suppresses gastric cancer cell biological activities[J]. Food Sci Nutr, 2020, 9(1):123-134.
19	Jin XZ, Qiao LN, Fan H, et al. Long non-coding RNA MSC-AS1 facilitates the proliferation and glycolysis of gastric cancer cells by regulating PFKFB3 expression[J]. Int J Med Sci, 2021, 18(1):546-554.
20	He ZZ, Long J, Yang C, et al. LncRNA DGCR5 plays a tumor-suppressive role in glioma via the miR-21/Smad7 and miR-23a/PTEN axes[J]. Aging (Albany NY), 2020, 12(20):20285-20307.
21	Li XJ, Zhou SX, Fan TY, et al. lncRNA DGCR 5/miR-27a-3p/BNIP3 promotes cell apoptosis in pancreatic cancer by regulating the p38 MAPK pathway[J]. Int J Mol Med, 2020, 46(2):729-739.
22	Huang HS, Yang XM, Chen J, et al. lncRNA DGCR5 inhibits the proliferation of colorectal cancer cells by downregulating miR-21[J]. Oncol Lett, 2019, 18(3):3331-3336.
23	Chen FK, Yin ST, Zhu JL, et al. lncRNA DGCR5 acts as a tumor suppressor in papillary thyroid carcinoma via sequestering miR-2861[J]. Exp Ther Med, 2019, 17(1):895-900.
24	Kang M F, Shi J Q, Li B H, et al. LncRNA DGCR5 regulates the non-small cell lung cancer cell growth, migration, and invasion through regulating miR-211-5p/EPHB6 axis[J]. Biofactors, 2019, 45(5):788-794.
25	Liu SL, Chu BF, Cai C, et al. DGCR5 promotes gallbladder cancer by sponging MiR-3619-5p via MEK/ERK1/2 and JNK/p38 MAPK pathways[J]. J Cancer, 2020, 11(18):5466-5477.
26	Jiang DQ, Wang C, He JJ. Long non-coding RNA DGCR5 incudes tumorigenesis of triple-negative breast cancer by affecting Wnt/β-catenin signaling pathway[J]. J BUON, 2020, 25(2):702-708.
27	Luo J, Zhu H, Jiang H, et al. The effects of aberrant expression of LncRNA DGCR5/miR-873-5p/TUSC3 in lung cancer cell progression[J]. Cancer Med, 2018, 7(7):3331-3341.
28	Liu S, Chu B, Cai C, et al. DGCR5 promotes gallbladder cancer by sponging miR-3619-5p via MEK/ERK1/2 and JNK/p38 MAPK pathways[J]. J Cancer, 2020, 11(18):5466-5477.
29	籍玉青, 尹永波, 邱金霞. lncRNA DGCR5通过靶向调控miR-21抑制鼻咽癌细胞的增殖、侵袭和迁移[J]. 中国老年学杂志, 2022, 42(11):2791-2795.
30	Liu SL, Cai C, Yang ZY, et al. DGCR5 is activated by PAX5 and promotes pancreatic cancer via targeting miR-3163/TOP2A and activating Wnt/β-catenin pathway[J]. Int J Biol Sci, 2021, 17(2):498-513.
31	Duan Y, Jia Y, Wang J, et al. Long noncoding RNA DGCR5 involves in tumorigenesis of esophageal squamous cell carcinoma via SRSF1-mediated alternative splicing of Mcl-1[J]. Cell Death Dis, 2021, 12(6):587. doi: 10.1038/s41419-021-03858-7.
32	Sun Y, Sun H. Propofol exerts anticancer activity on hepatocellular carcinoma cells by raising lncRNA DGCR5[J]. J Cell Physiol, 2020, 235(3):2963-2972.

基因	序列（5'→3'）	预期扩增产物长度（bp）
lncRNA DGCR5	上游CACGAGTGTAGTGCCCAGTT	75
	下游GGTCAGGGACCTTTGTCGTT
β-actin	上游AAGTACTCCGTGTGGATCGG	136
	下游ATGCTATCACCTCCCCTGTG

基因	序列（5'→3'）	预期扩增产物长度（bp）
lncRNA DGCR5	上游CACGAGTGTAGTGCCCAGTT	75
	下游GGTCAGGGACCTTTGTCGTT
β-actin	上游AAGTACTCCGTGTGGATCGG	136
	下游ATGCTATCACCTCCCCTGTG

分组	A值	克隆数（个）
对照组	1.14 ± 0.10	163.45 ± 9.87
低剂量组	0.98 ± 0.08^a	121.63 ± 8.43^a
中剂量组	0.77 ± 0.06^ab	93.87 ± 6.96^ab
高剂量组	0.60 ± 0.05^abc	72.65 ± 5.68^abc
F值	89.533	222.182
P值	< 0.001	< 0.001

分组	A值	克隆数（个）
对照组	1.14 ± 0.10	163.45 ± 9.87
低剂量组	0.98 ± 0.08^a	121.63 ± 8.43^a
中剂量组	0.77 ± 0.06^ab	93.87 ± 6.96^ab
高剂量组	0.60 ± 0.05^abc	72.65 ± 5.68^abc
F值	89.533	222.182
P值	< 0.001	< 0.001

分组	MMP2	MMP9	细胞迁移数量（个）	细胞侵袭数量（个）	划痕愈合率（%）
对照组	0.85 ± 0.07	0.79 ± 0.06	243.49 ± 15.42	187.36 ± 13.24	98.74 ± 2.56
低剂量组	0.69 ± 0.05^a	0.71 ± 0.07^a	216.73 ± 13.16^a	153.41 ± 9.16^a	86.43 ± 5.14^a
中剂量组	0.49 ± 0.03^ab	0.58 ± 0.04^ab	173.42 ± 12.04^ab	116.98 ± 6.13^ab	72.59 ± 4.71^ab
高剂量组	0.37 ± 0.02^abc	0.41 ± 0.03^abc	136.09 ± 8.39^abc	84.26 ± 3.79^abc	57.62 ± 2.93^abc
F/F'值	F' = 187.034	90.191	129.006	F' = 230.585	177.522
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

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