Chloroprocaine inhibits the growth and metastasis of liver cancer Huh-7 cells in vitro by regulating the expression of circRNA-ZKSCAN1

doi:10.3877/cma.j.issn.2095-1221.2024.04.004

Abstract

Abstract:

Objective

To investigate the effect of chloroprocaine (CP) on the biological behavior of human hepatoma cell Huh-7 and its regulation on circZKSCAN1.

Methods

Huh-7 cells were treated with different doses of CP, and subsequent experimental treatments: pcDNA and pcDNA-circZKSCAN1 were transfected into Huh-7 cells, respectively. si-NC and si-circZKSCAN1 were transfected into Huh-7 cells and treated with CP (10 mmol/L). QRT-PCR detected the expression of circZKSCAN1. CCK-8, plate clone formation assay, scratch assay, and Transwell chamber were used to detect cell proliferation, clone formation, migration and invasion abilities, respectively. Western blot was used to detect the expression of E-cadherin and N-cadherin. T test was used for comparison between the two groups, One-way analysis of variance was used for statistical comparison among multiple groups, and the SNK-q method was used for further pairwise statistical comparison.

Results

After the treatment of CP, the inhibition rate of cell proliferation [ (24.20 ± 2.21) %, (48.25 ± 4.42) %, (65.19 ± 5.49) % vs (0.00 ± 0.00) %], the protein level of E-cadherin, and the expression of circZKSCAN1 [1.51 ± 0.13, 2.11 ± 0.22, 2.92 ± 0.27 vs 1.00 ± 0.00] were increased (P < 0.05), while the number of cell clones and invasive cells [ (99.96 ± 7.03), (79.87 ± 4.96), (58.64 ± 3.99) vs (124.01 ± 10.98) pcs] was decreased (P < 0.05), and the wound healing rate [ (53.71 ± 4.19) %, (40.61 ± 4.58) %, (24.13 ± 2.22) % vs (67.15 ± 5.17) %], the protein level of N-cadherin were decreased (P < 0.05), and they were dose-dependent. After transfection of pcDNA-circZKSCAN1, the inhibition rate of cell proliferation [ (57.19 ± 5.27) % vs (7.13 ± 0.54) %] and the protein level of E-cadherin were higher than those of transfected pcDNA (P < 0.05), while the number of clones and invasive cells [ (31.11 ± 3.01) % vs (68.07 ± 4.76) %] was lower than those of transfected pcDNA (P < 0.05), and the wound healing rate [ (66.33 ± 5.84) vs (126.12 ± 11.74) pcs], the protein level of N-cadherin was lower than that of transfected pcDNA (P < 0.05). Transfection of si-circZKSCAN1 could reduce the effect of CP on the biological behavior of Huh-7 cells.

Conclusion

CP could inhibit the growth and metastasis of liver cancer Huh-7 cells by up-regulating the expression of circZKSCAN1.

Key words: Chloroprocaine, circZKSCAN1, Liver cancer, Cell proliferation, Migration, Invasion

Yuli Zeng, Farong Lei, Hui Xiao, Deliang Qiu, Jing Xie, Xun Wu. Chloroprocaine inhibits the growth and metastasis of liver cancer Huh-7 cells in vitro by regulating the expression of circRNA-ZKSCAN1[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(04): 220-228.

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Figures/Tables 15

References 23

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基因	引物序列（5'→3'）	预期扩增长度（bp）
circRNA-ZKSCAN1	上游GGAATACCGCCCCGATAGTG	226
	下游TCCAAACAGGGTCTGTGCTC
GAPDH	上游GGAGCGAGATCCCTCCAAAAT	198
	下游GGCTGTTGTCATACTTCTCATGG

基因	引物序列（5'→3'）	预期扩增长度（bp）
circRNA-ZKSCAN1	上游GGAATACCGCCCCGATAGTG	226
	下游TCCAAACAGGGTCTGTGCTC
GAPDH	上游GGAGCGAGATCCCTCCAAAAT	198
	下游GGCTGTTGTCATACTTCTCATGG

分组	实验次数	抑制率（%）	细胞克隆形成数（个）
对照组	3	0.00 ± 0.00	95.67 ± 7.79
CP低剂量组	3	24.20 ± 2.21^a	74.58 ± 6.09^a
CP中剂量组	3	48.25 ± 4.42^ab	58.94 ± 4.13^ab
CP高剂量组	3	65.19 ± 5.49^abc	38.58 ± 3.41^abc
F/H值		H = 177.95	F = 55.42
P值		< 0.001	< 0.001

分组	实验次数	抑制率（%）	细胞克隆形成数（个）
对照组	3	0.00 ± 0.00	95.67 ± 7.79
CP低剂量组	3	24.20 ± 2.21^a	74.58 ± 6.09^a
CP中剂量组	3	48.25 ± 4.42^ab	58.94 ± 4.13^ab
CP高剂量组	3	65.19 ± 5.49^abc	38.58 ± 3.41^abc
F/H值		H = 177.95	F = 55.42
P值		< 0.001	< 0.001

分组	实验次数	划痕愈合率（%）	侵袭细胞数（个）	E-cadherin蛋白	N-cadherin蛋白
对照组	3	67.15 ± 5.17	124.01 ± 10.98	0.28 ± 0.02	0.64 ± 0.05
CP低剂量组	3	53.71 ± 4.19^a	99.96 ± 7.03^a	0.41 ± 0.04^a	0.52 ± 0.04^a
CP中剂量组	3	40.61 ± 4.58^ab	79.87 ± 4.96^ab	0.55 ± 0.05^ab	0.38 ± 0.04^ab
CP高剂量组	3	24.13 ± 2.22^abc	58.64 ± 3.99^abc	0.73 ± 0.06^abc	0.23 ± 0.03^abc
F值		57.76	44.43	58.01	57.02
P值		< 0.001	< 0.001	< 0.001	< 0.001

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