Dual phase modeling of Chrysotile carcinogenesis from 3D cell transformation to orthotopic tumors

Our plain-English summary of Dual phase modeling of Chrysotile carcinogenesis from 3D cell transformation to orthotopic tumors, by Y. Gao, W. Yu, R. Li, et al (Scientific Reports, 2026).

The study

• Authors: Y. Gao, W. Yu, R. Li, Q. Wang, M. Zhang, Y. Zhong, H. Xia, F. Zhang, L. Zhu
• Journal: Scientific Reports
• Published: 2026;16(1):2199
• Read it: PubMed · Publisher · Free full text

Abstract

Chrysotile asbestos, a well-established carcinogen, is known to induce various malignant cancers, contributing to an estimated annual death toll of 107,000 individuals due to asbestos-related diseases. While cell and animal models play a crucial role in elucidating the carcinogenic mechanisms of asbestos, existing models are plagued by prolonged experimental durations. In this investigation, we established a novel NIH/3T3 cell model of asbestos-induced malignant transformation using 3D culture techniques, followed by the development of a corresponding mouse model via orthotopic transplantation. Subsequent administration of the HIF-1α inhibitor PX-478 to both models allowed for the assessment of model reliability through the observation of malignant phenotypes and associated protein alterations.

© 2026. The Author(s).

In plain English

This laboratory study set out to build a faster model of chrysotile carcinogenesis. Mouse embryonic fibroblast NIH/3T3 cells were grown in a three-dimensional matrix gel and exposed three times, each for 48 hours, over 21 days, to UICC standard chrysotile at 0.625 micrograms per square centimetre; at that dose short-term proliferation was 94.2% in 2D and 97.2% in 3D. The chrysotile-exposed 3D cells (3D-Asb) formed more soft-agar colonies, showed less apoptosis, more cells in G1, and greater wound healing, migration and invasion than control cells; proliferation did not change significantly. These transformed cells, fluorescently labelled, were injected into nude mouse lungs and produced lung tumours adhering to the chest wall within four weeks. In cells, the HIF-1 alpha inhibitor PX-478 reduced proliferation, migration and invasion and lowered TWIST protein, with no significant change in N-cadherin or vimentin; in the mice, lung fluorescence was significantly lower in the PX-478 group on days 14, 21 and 28, while reductions in lung TWIST and vimentin were not statistically significant. The authors concluded the dual model is reliable and shortens modelling time.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory experimental study: in vitro 3D cell transformation model plus orthotopic transplantation animal experiment in nude mice, with drug (PX-478) testing in both
• Who was studied: NIH/3T3 mouse embryonic fibroblast cell line and nude mice; in the PX-478 experiment 12 mice in the control group and 12 in the experimental group, Hangzhou Medical College, China
• Exposure: UICC standard chrysotile asbestos, repeated 48-hour exposures at 0.625 micrograms per square centimetre in 3D culture over 21 days; also PX-478 (HIF-1 alpha inhibitor) at 5, 10 and 20 micromolar in cells and injected in mice
• Main results:
  • Short-term chrysotile toxicity at 48 hours: cell proliferation was 94.2% in 2D and 97.2% in 3D at 0.625 micrograms per square centimetre, and 86.4% in 2D and 93.4% in 3D at 1.25 micrograms per square centimetre
  • The chrysotile-exposed 3D group (3D-Asb) showed anchorage-independent growth with a higher clone formation rate than the 3D control group (P less than 0.01)
  • 3D-Asb cells had decreased apoptosis versus controls (P less than 0.01), and no significant change in proliferation
  • A larger proportion of 3D-Asb cells was blocked in G1 phase, with a shortened S + M proportion, compared with control cells
  • 3D-Asb cells showed greater wound healing (P less than 0.01), Transwell migration (P less than 0.01) and Transwell invasion (P less than 0.001) than 3D-CT cells
  • In the first nude mouse model, five mice received intrapulmonary injection of the chrysotile-transformed cells
  • In that first model, lung fluorescence was visible on day 7 and increased on days 14 and 21
  • One mouse died after the third imaging session, and the remaining mice were euthanized
  • Lung tissue of those mice showed chest-wall adhesions and irregular tumours in both lungs
  • In 3D-Asb cells, PX-478 at 20 micromolar reduced proliferation at 24 hours (P less than 0.05), and 10 and 20 micromolar reduced it at 48 hours (P less than 0.001)
  • Transwell invasion by 3D-Asb cells fell at PX-478 concentrations of 5, 10 and 20 micromolar compared with control (P less than 0.001), decreasing with higher concentration
  • In treated cells, TWIST protein decreased in the 20 micromolar PX-478 group while N-cadherin remained relatively stable
  • Vimentin protein in the malignantly transformed cells showed no significant change after 24 and 48 hours of PX-478 treatment
  • In the PX-478 mouse experiment, there was no significant difference in fluorescence between the two groups on day 7
  • On day 14, lung fluorescence intensity in the PX-478 group was lower than the control group, a statistically significant difference
  • On day 21, lung fluorescence intensity in the PX-478 group was lower than the control group, a statistically significant difference
  • On day 28, lung fluorescence intensity in the PX-478 group was lower than the control group, a statistically significant difference
  • Body weight changes in the two mouse groups were not obvious
  • In mouse lung tissue, N-cadherin showed no significant change, and the downward trends in TWIST and vimentin were not statistically significant
• The authors' conclusion: The authors concluded that the 3D cell model and the fluorescently labelled nude mouse orthotopic model were reliable, took less time to build, allowed real-time observation, and could serve as a platform for studying asbestos carcinogenic mechanisms and other toxins
• Limitations the authors state:
  • The authors noted that the PX-478 effect on TWIST was not statistically significant in the mouse model
  • The authors noted that despite the reduction in TWIST, N-cadherin and vimentin levels did not change in either model

Topics

The National Library of Medicine indexes this study under subjects our site covers: Chrysotile.

Medical terms in this study, explained in our Lexicon: Chrysotile, Carcinogenesis, Carcinogenic, Malignant Transformation.

Related studies

• Continuous exposure to chrysotile asbestos can cause transformation of human mesothelial cells via HMGB1 and TNF-α signaling — F. Qi, G. Okimoto, et al. (2013)
• Malignant transformation of immortalized human bronchial epithelial cells by asbestos fibers — T.K. Hei, L.J. Wu, et al. (1997)
• In vitro transformation of rat pleural mesothelial cells by chrysotile fibres and/or benzo[a]pyrene — M.J. Paterour, J. Bignon, et al. (1985)
• Asbestos promotes morphological transformation and elevates expression of a gene family invariably induced by tumor promoters in C3H/10T1/2 cells — C.L. Parfett, R. Pilon, et al. (1996)
• Dual preventive benefits of iron elimination by desferal in asbestos-induced mesothelial carcinogenesis — L. Jiang, S.H. Chew, et al. (2016)
• Generation of human embryonic stem cell models to exploit the EWSR1-CREB fusion promiscuity as a common pathway of transformation in human tumors — F. Vanoli, B. Meskauskaite, et al. (2021)

Browse more research in Scientific Reports, or return to medical research.