Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression

Abstract Background Liver cancer stem cells (LCSCs) are critical drivers of metastasis and chemoresistance in hepatocellular carcinoma (HCC). Proline-rich tyrosine kinase 2 (Pyk2) has been implicated in tumor progression, but its role in LCSC stemness and HCC malignancy remains unclear. This study e...

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Main Authors: Meng Sha, Jiang Zhang, Jin-kai Liu, Xiao-ye Qu, Chuan Shen, Ying Tong, Jie Cao
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Cancer Cell International
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Online Access:https://doi.org/10.1186/s12935-025-03841-8
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author Meng Sha
Jiang Zhang
Jin-kai Liu
Xiao-ye Qu
Chuan Shen
Ying Tong
Jie Cao
author_facet Meng Sha
Jiang Zhang
Jin-kai Liu
Xiao-ye Qu
Chuan Shen
Ying Tong
Jie Cao
author_sort Meng Sha
collection DOAJ
description Abstract Background Liver cancer stem cells (LCSCs) are critical drivers of metastasis and chemoresistance in hepatocellular carcinoma (HCC). Proline-rich tyrosine kinase 2 (Pyk2) has been implicated in tumor progression, but its role in LCSC stemness and HCC malignancy remains unclear. This study explores the effects of Pyk2 and its regulation by miR-23b-3p on LCSC function and HCC progression. Methods LCSCs were enriched from HepG2 and HCCLM3 cell lines, and Pyk2 knockdown was induced through siRNA transfection, with or without miR-23b-3p inhibitor co-transfection. We assessed cell proliferation, sphere formation, migration, invasion, and chemosensitivity. Stemness markers (Nanog, Oct4, Sox2, KLF4, and Bmi1) and Pyk2/Src signaling were analyzed via RT-qPCR, Western blotting, and immunohistochemistry. In vivo, tumor growth and Pyk2/Src expressions were evaluated in a BALB/c mouse xenograft model. Results Pyk2 expression was significantly elevated in the identified LCSCs compared to the parental HCCs. Pyk2 knockdown significantly suppressed the LCSCs proliferation, sphere formation, migration, invasion, and enhanced chemosensitivity. The expression of stemness markers and miR-23b-3p was significantly inhibited in HCCLM3-LCSCsiPyk2 cells. miR-23b-3p inhibition restored Pyk2 level and Src phosphorylation, reversing the suppressive effects of Pyk2 knockdown. In BALB/c mice, tumor volume, weight, and Pyk2/Src expressions were significantly elevated in HCCLM3-LCSC and HCCLM3-LCSCsiPyk2+miR-23b-3p inhibitor groups comparing to HCCLM3/HCCLM3-LCSCsiPyk2 groups, whereas were even heightened in the HCCLM3-LCSC + miR-23b-3p inhibitor group. Conclusions Inhibiting Pyk2/Src expression by miR-23b-3p suppressed LCSCs function and aggravated HCC progression. Trial registration Not applicable.
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spelling doaj-art-ee19f2e789b84220a9e7e2d4991c014b2025-08-20T02:30:59ZengBMCCancer Cell International1475-28672025-06-0125111510.1186/s12935-025-03841-8Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progressionMeng Sha0Jiang Zhang1Jin-kai Liu2Xiao-ye Qu3Chuan Shen4Ying Tong5Jie Cao6Department of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Liver Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityAbstract Background Liver cancer stem cells (LCSCs) are critical drivers of metastasis and chemoresistance in hepatocellular carcinoma (HCC). Proline-rich tyrosine kinase 2 (Pyk2) has been implicated in tumor progression, but its role in LCSC stemness and HCC malignancy remains unclear. This study explores the effects of Pyk2 and its regulation by miR-23b-3p on LCSC function and HCC progression. Methods LCSCs were enriched from HepG2 and HCCLM3 cell lines, and Pyk2 knockdown was induced through siRNA transfection, with or without miR-23b-3p inhibitor co-transfection. We assessed cell proliferation, sphere formation, migration, invasion, and chemosensitivity. Stemness markers (Nanog, Oct4, Sox2, KLF4, and Bmi1) and Pyk2/Src signaling were analyzed via RT-qPCR, Western blotting, and immunohistochemistry. In vivo, tumor growth and Pyk2/Src expressions were evaluated in a BALB/c mouse xenograft model. Results Pyk2 expression was significantly elevated in the identified LCSCs compared to the parental HCCs. Pyk2 knockdown significantly suppressed the LCSCs proliferation, sphere formation, migration, invasion, and enhanced chemosensitivity. The expression of stemness markers and miR-23b-3p was significantly inhibited in HCCLM3-LCSCsiPyk2 cells. miR-23b-3p inhibition restored Pyk2 level and Src phosphorylation, reversing the suppressive effects of Pyk2 knockdown. In BALB/c mice, tumor volume, weight, and Pyk2/Src expressions were significantly elevated in HCCLM3-LCSC and HCCLM3-LCSCsiPyk2+miR-23b-3p inhibitor groups comparing to HCCLM3/HCCLM3-LCSCsiPyk2 groups, whereas were even heightened in the HCCLM3-LCSC + miR-23b-3p inhibitor group. Conclusions Inhibiting Pyk2/Src expression by miR-23b-3p suppressed LCSCs function and aggravated HCC progression. Trial registration Not applicable.https://doi.org/10.1186/s12935-025-03841-8Hepatocellular carcinomaLiver cancer stem cellsProline-rich tyrosine kinase 2miR-23b-3pSrc
spellingShingle Meng Sha
Jiang Zhang
Jin-kai Liu
Xiao-ye Qu
Chuan Shen
Ying Tong
Jie Cao
Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
Cancer Cell International
Hepatocellular carcinoma
Liver cancer stem cells
Proline-rich tyrosine kinase 2
miR-23b-3p
Src
title Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
title_full Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
title_fullStr Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
title_full_unstemmed Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
title_short Inhibiting Pyk2/Src expression by miR-23b-3p suppressed liver cancer stem cell function and hepatic carcinoma progression
title_sort inhibiting pyk2 src expression by mir 23b 3p suppressed liver cancer stem cell function and hepatic carcinoma progression
topic Hepatocellular carcinoma
Liver cancer stem cells
Proline-rich tyrosine kinase 2
miR-23b-3p
Src
url https://doi.org/10.1186/s12935-025-03841-8
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