Inhibitory effect and mechanisms of psoralen on proliferation and migration of bladder cancer T24 cells

doi:10.3877/cma.j.issn.2095-1221.2022.04.001

Abstract

Abstract:

Objective

To investigate the effects of psoralen on the viability, cell cycle, apoptosis, and migration of human bladder cancer T24 cells and its mechanisms.

Methods

Bladder cancer cells were treated with cell culture medium, 3‰ dimethyl sulfoxide (DMSO) and different concentrations of psoralen and divided into normal control group, DMSO group and psoralen groups (10、30、50、100 μg/mL) . CCK-8 was used to detect cell viabilities. FACS was performed to detect cell cycles and apoptosis. Scratch tests were used to detecte the scratch healing rates. The expression levels of phosphatidylinositol-3-kinase (PI3K) and protein kinase B (AKT) mRNA were detected by RT-qPCR, and the expression and phosphorylation levels of PI3K and AKT protein were detected by Western blot. One-way ANOVA was used for comparison among multiple groups, and LSD-t test was used for comparison between groups.

Results

Compared with DMSO groups, the cell viabilities were decreased gradually with the increase of psoralen concentration and action time (P < 0.05) , except the cells were treated with 10 μg/mL psoralen for 24 h. Compared with DMSO group, the proportion of cells in G1 phase was increased after treatment with 30, 100 μg/mL psoralen for 24 h. The proportion of cells in G2/M phase was decreased, the apoptosis rates were increased [ (9.16±0.97) %、(15.45±1.57) % vs (1.02±0.36) %], the scratch healing rates were decreased [24 h: (45.00±3.44) %, (27.60± 2.21) %vs (66.10±2.61) %, 48 h: (70.00± 3.40) %, (45.17±2.44) %vs (85.17± 3.85) %], PI3K and AKT mRNA expression levels, protein expression levels and phosphorylation levels were all decreased (all P < 0.05) .

Conclusion

Psoralen decreased cell viability, blocked cell cycle, induced apoptosis and inhibited cell migration. Its mechanism may be that psoralen down-regulates the expression levels of PI3K and AKT mRNA and the expression levels and phosphorylation levels of PI3K and AKT protein.

Key words: Psoralen, Antitumor, Bladder cancer, PI3K, AKT

Mingfang Weng, Rong Liu, Junjie Wei, Qin Lin, Xinghui Sun, dong Wang, Weizhen Wu. Inhibitory effect and mechanisms of psoralen on proliferation and migration of bladder cancer T24 cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2022, 12(04): 193-199.

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Figures/Tables 10

References 21

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基因	序列（5'- 3'）	预期扩增产物长度（bp）
PI3K	上游ACCACTACCGGAATGAATCTCT	207
	下游GGGATGTGCGGGTATATTCTTC
AKT	上游CACAGGTCGCTACTATGCC	152
	下游GGTCGTGGGTCTGGAATG
β-actin	上游CATGTACGTTGCTATCCAGGC	250
	下游CTCCTTAATGTCACGCACGAT

基因	序列（5'- 3'）	预期扩增产物长度（bp）
PI3K	上游ACCACTACCGGAATGAATCTCT	207
	下游GGGATGTGCGGGTATATTCTTC
AKT	上游CACAGGTCGCTACTATGCC	152
	下游GGTCGTGGGTCTGGAATG
β-actin	上游CATGTACGTTGCTATCCAGGC	250
	下游CTCCTTAATGTCACGCACGAT

分组	实验次数	24 h	48 h	72 h	96 h	F值	P值
正常对照	3	99.71±4.70	103.32±2.05	102.65±4.61	102.24±0.99
DMSO	3	100.00±0.00	100.00±0.00	100.00±0.00	100.00±0.00
10 µg/mL	3	96.50±2.70	94.53±1.68^a	90.34±3.07^aef	85.50±2.29^aefg	19.062	< 0.001
30 µg/mL	3	93.64±1.67^a	87.29±2.72^abe	83.67±5.76^abe	75.82±2.09^abefg	26.956	< 0.001
50 µg/mL	3	89.72±2.80^abc	83.47±2.12^abce	73.56±2.34^abcef	64.95±4.19^abcefg	79.297	< 0.001
100 µg/mL	3	85.45±2.21^abcd	77.60±2.45^abcde	64.24±3.00^abcdef	55.45±3.06^abcdefg	157.463	< 0.001
F值		21.192	56.974	51.392	124.302
P值		< 0.001	< 0.001	< 0.001	< 0.001

分组	实验次数	24 h	48 h	72 h	96 h	F值	P值
正常对照	3	99.71±4.70	103.32±2.05	102.65±4.61	102.24±0.99
DMSO	3	100.00±0.00	100.00±0.00	100.00±0.00	100.00±0.00
10 µg/mL	3	96.50±2.70	94.53±1.68^a	90.34±3.07^aef	85.50±2.29^aefg	19.062	< 0.001
30 µg/mL	3	93.64±1.67^a	87.29±2.72^abe	83.67±5.76^abe	75.82±2.09^abefg	26.956	< 0.001
50 µg/mL	3	89.72±2.80^abc	83.47±2.12^abce	73.56±2.34^abcef	64.95±4.19^abcefg	79.297	< 0.001
100 µg/mL	3	85.45±2.21^abcd	77.60±2.45^abcde	64.24±3.00^abcdef	55.45±3.06^abcdefg	157.463	< 0.001
F值		21.192	56.974	51.392	124.302
P值		< 0.001	< 0.001	< 0.001	< 0.001

分组	实验次数	G1	S	G2/M
DMSO	3	57.91±0.84	25.73±0.17	16.37±1.00
30 µg/mL	3	62.34±1.13^a	32.77±1.35^a	4.89±1.38^a
100 µg/mL	3	72.08±2.75^ab	25.56±0.83^b	2.36±1.97^a
F值		49.678	59.844	73.895
P值		< 0.001	< 0.001	< 0.001

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