5-氟尿嘧啶联合重组人细胞珠蛋白治疗缺氧诱导化疗耐受肝细胞癌的体外实验

doi:10.3969/j.issn.1006-5725.2024.22.005

Abstract

Abstract:

Objective This study explores the efficacy and underlying mechanisms of combining rhCYGB with 5-FU to target hypoxia-induced treatment resistance in liver cancer. The aim is to develop a novel combinatorial therapy strategy for improving outcomes in patients with refractory liver cancer. Methods The half-maximal inhibitory concentration （IC₅₀） of drugs on liver cancer cells under normoxia and hypoxia was determined， and dose-response curves were generated to assess sensitivity to 5-FU. The combined effects of rhCYGB and 5-FU were analyzed withCompuSyn and SynergyFinder 3.0. Tumor stem cell sphere formation assays and flow cytometry for CD133-positive cells were conducted to evaluate the impact on cancer stemness. Wound healing assays assessed the effects on migration. Results The IC₅₀ values under hypoxia exhibited a significant fold change compared to normoxia（P < 0.05）， specifically a 15.27-fold， 4.25-fold， and 2.34-fold increase for Hep3B， Huh7， and HepG2 cells respectively. Assessment of drug combination effects demonstrated a synergistic interaction between 5-FU and rhCYGB. Compared to the 5-FU monotherapy group， the combination of 5-FU and rhCYGB exerted an inhibitory effect on the formation of liver cancer stem cell spheres （P < 0.05） and significantly downregulated the proportion of CD133-positive subpopulations in Hep3B cells （P < 0.05）. Wound healing assay results revealed a synergistic inhibitory effect on the migration of Hep3B cells after 48 hours of treatment with rhCYGB combined with 5-FU undernormoxia （P < 0.05）. Conclusions The combination of rhCYGB and 5-FU demonstrates synergistic effects in liver cancer. The underlying mechanism may involve the modulation of stemness and cell migration capacity.

Key words: rhCYGB, hypoxia, combination strategy, cancer stem cell, hepatocellular carcinoma

CLC Number:

R735.7

Yi WANG,Shuo HE,Siyun YANG,Jun. ZHANG. In vitro study of 5⁃FU combined with rhCYGB in treating hypoxia⁃induced chemotherapy resistance of hepatocellular carcinoma[J]. The Journal of Practical Medicine, 2024, 40(22): 3146-3154.

Figures/Tables 9

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References 25

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细胞系	缺氧	常氧	耐药比值	P值
细胞系	IC₅₀/（μmol/L）	IC₅₀/（μmol/L）	耐药比值	P值
Hep3B	454.20 ± 42.44	29.75 ± 2.48	15.27	< 0.01
Huh7	73.58 ± 1.71	17.30 ± 0.52	4.25	< 0.05
HepG2	45.40 ± 2.61	19.36 ± 0.76	2.34	< 0.05

rhCYGB	5-FU	Hep3B		Huh7		HepG2
rhCYGB	5-FU	常氧	缺氧	常氧	缺氧	常氧	缺氧
10 nmol/L	20 μmol/L	0.80 ± 0.02	3.43 ± 3.16	0.51 ± 0.00	0.36 ± 0.03	1.45 ± 0.32	1.94 ± 0.21
10 nmol/L	40 μmol/L	1.17 ± 0.16	0.99 ± 0.01	0.81 ± 0.02	0.51 ± 0.11	1.34 ± 0.04	1.81 ± 0.77
10 nmol/L	80 μmol/L	0.43 ± 0.00	0.30 ± 0.14	1.14 ± 0.00	0.98 ± 0.36	0.63 ± 0.00	0.55 ± 0.56
25 nmol/L	20 μmol/L	0.86 ± 0.07	0.31 ± 0.15	0.65 ± 0.00	0.65 ± 0.01	0.75 ± 0.05	1.07 ± 0.37
25 nmol/L	40 μmol/L	1.49 ± 0.04	0.30 ± 0.27	0.98 ± 0.07	0.74 ± 0.01	0.81 ± 0.00	2.11 ± 0.50
25 nmol/L	80 μmol/L	1.21 ± 0.10	0.81 ± 0.64	1.37 ± 0.00	1.04 ± 0.01	1.00 ± 0.01	0.25 ± 0.04
50 nmol/L	20 μmol/L	0.56 ± 0.04	0.51 ± 0.40	1.07 ± 0.00	1.25 ± 0.03	1.32 ± 0.01	0.24 ± 0.00
50 nmol/L	40 μmol/L	0.96 ± 0.03	0.19 ± 0.00	1.30 ± 0.00	1.33 ± 0.04	1.42 ± 0.01	0.27 ± 0.00
50 nmol/L	80 μmol/L	0.43 ± 0.01	0.23 ± 0.00	1.78 ± 0.01	1.56 ± 0.12	1.60 ± 0.01	0.33 ± 0.00

组别	常氧
	愈合率/%		P值
	48 h	72 h	vs.对照组		vs.联合组
	48 h	72 h	48 h	72 h	48 h	72 h
对照组	41.66 ± 2.29	63.16 ± 2.07
rhCYGB组	44.38 ± 1.80	62.07 ± 3.15	0.231	0.659	< 0.05	< 0.05
5-FU组	34.65 ± 2.55	42.58 ± 4.61	< 0.05	< 0.001	< 0.05	< 0.05
联合组	28.41 ± 2.88	33.47 ± 1.62
组别	缺氧
	愈合率/%		P值
	48 h	72 h	vs.对照组		vs.联合组
	48 h	72 h	48 h	72 h	48 h	72 h
对照组	45.63 ± 2.73	45.63 ± 2.73
rhCYGB组	37.23 ± 2.39	51.27 ± 5.18	< 0.01	0.425	0.516	0.180
5-FU组	40.29 ± 1.90	49.41 ± 2.11	< 0.05	< 0.05	0.134	0.222
联合组	37.33 ± 2.70	45.15 ± 4.68

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In vitro study of 5⁃FU combined with rhCYGB in treating hypoxia⁃induced chemotherapy resistance of hepatocellular carcinoma

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细胞	5-FU浓度/（μmol/L）
Hep3B	0.01	0.1	1	5	10	20	40	80	100	200	400	1 000
Huh7	0.01	1	5	10	20	40	80	125	250	500	1 000	2 000
HepG2	0.01	1	5	10	20	40	80	125	250	500	1 000	2 000