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Article Open Access

Hydroxypropyl‑β‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑κB‑mediated ferroptosis

  • Authors:
    • Wei-Wei Zheng
    • Jia-Xuan Ding
    • Yang-Xin Liang
    • Ling Ye
  • View Affiliations / Copyright

    Affiliations: Department of Radiation Oncology, Ma'anshan People's Hospital, Ma'anshan, Anhui 243000, P.R. China, Department of Radiation Oncology, Sun Yat‑sen Memorial Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510120, P.R. China
    Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 157
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    Published online on: September 16, 2025
       https://doi.org/10.3892/or.2025.8990
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Abstract

Thymoquinone (TQ) is a quinone isolated from the black seed Nigella sativa and has been extensively investigated in the pharmaceutical field due to its promising therapeutic value. There have been reports on the potential anti‑cancer properties of TQ, whereas the clinical application of TQ is greatly restricted by its low water solubility and poor delivery. In the present study, TQ was encapsulated into hydroxypropyl‑β‑cyclodextrin (HP‑β‑CD) using the freeze‑drying method to form a TQ/HP‑β‑CD inclusion complex. Then, the TQ/HP‑β‑CD inclusion complex was characterized by Fourier transform infrared, differential scanning calorimetry, X‑ray diffraction and scanning electron microscopy. HP‑β‑CD as an excipient significantly enhances the water solubility of TQ, and TQ/HP‑β‑CD demonstrated excellent biocompatibility on normal cells both in vitro and in vivo. Moreover, the complexation with HP‑β‑CD enhanced the anticancer activity of TQ against non‑small cell lung cancer cells and its mechanism of action is related to ferroptosis mediated by NF‑κB. Such an enhanced cytotoxic effect may be attributed to the effective complexation of TQ in HP‑β‑CD, which enhances its water solubility and bioavailability.
View Figures

Figure 1

Schematic illustration of TQ/HP-β-CD
inclusion complex. TQ, thymoquinone; HP-β-CD,
hydroxypropyl-β-cyclodextrin

Figure 2

Characterization of TQ/HP-β-CD
inclusion complex. (A) The Fourier transform infrared spectra, (B)
X-ray diffraction diagram, (C) Differential scanning calorimetry
and (D) scanning electron microscope images of TQ, HP-β-CD and
TQ/HP-β-CD inclusion complex. TQ, thymoquinone; HP-β-CD,
hydroxypropyl-β-cyclodextrin.

Figure 3

(A) Phase solubility diagram of
TQ/HP-β-CD host-guest system. (B) In vitro release curve of
free TQ and TQ/HP-β-CD inclusion complex in PBS (pH=7.4 at 37°C).
TQ, thymoquinone; HP-β-CD, hydroxypropyl-β-cyclodextrin

Figure 4

(A) Hemocompatibility of TQ/HP-β-CD
inclusion complex. Effect of different concentrations of TQ/HP-β-CD
inclusion complex on red blood cell hemolysis after 1 h and 24 h of
treatment. (B) Spectrophotometry was used to quantitatively
evaluate the degree of hemolysis in each group. TQ, thymoquinone;
HP-β-CD, hydroxypropyl-β-cyclodextrin.

Figure 5

(A) The growth inhibitory effect of
TQ/H2O, TQ/DMSO, HP-β-CD and TQ/HP-β-CD inclusion
complex on A549 and HCC827 cells was determined by Cell Counting
Kit-8 assay. (B) Cell proliferation capacity was detected by EdU
staining after treatment with TQ/H2O and TQ/HP-β-CD for
24 h in non-small-cell lung cancer cells. (C) The effects of
TQ/H2O and TQ/HP-β-CD on the sprout outgrowth of A549
and HCC827 cells. TQ, thymoquinone; HP-β-CD,
hydroxypropyl-β-cyclodextrin.

Figure 6

TQ/HP-β-CD induces ferroptosis in
NSCLC cells. The effect of TQ on ROS generation in NSCLC cells was
detected by (A) DHCA staining and (B) flow cytometry. (C) Detection
of lipid peroxidation level in NSCLC cells by C11-BODIPY 581/591
staining. (D) The levels of MDA, GSH and the ratio of GSH/GSSG in
NSCLC cells were detected by ELISA. *P<0.05 vs. control group.
TQ, thymoquinone; HP-β-CD, hydroxypropyl-β-cyclodextrin; NSCLC,
non-small-cell lung cancer; MDA, malondialdehyde; GSH, reduced
glutathione; GSSG, oxidized glutathione.

Figure 7

Ferroptosis-dependent effect of
TQ/HP-β-CD against NSCLC cells. Cell viability detection in A549
and HCC827 cells treated with or without Ferrostatin-1 in the
presence of TQ/HP-β-CD (A). Analysis of ROS level in A549 and
HCC827 cells treated with or without Ferrostatin-1 in the presence
of TQ/HP-β-CD (B). Detection of lipid peroxidation level in NSCLC
cells treated with or without Ferrostatin-1 in the presence of
TQ/HP-β-CD (C). The levels of MDA, GSH and the ratio of GSH/GSSG in
NSCLC cells were detected by ELISA. (D) *P<0.05 vs. control
group; #P<0.05 vs. TQ/HP-β-CD group. TQ,
thymoquinone; HP-β-CD, hydroxypropyl-β-cyclodextrin; NSCLC,
non-small-cell lung cancer; MDA, malondialdehyde; GSH, reduced
glutathione; GSSG, oxidized glutathione.

Figure 8

TQ/HP-β-CD induces ferroptosis in
non-small-cell lung cancer cells by inhibiting NF-κB activity. (A)
Electrophoretic mobility shift assay showing NF-κB DNA binding
activity in A549 cells. (B) Detection of lipid peroxidation level
in A549 cells. (C) Cell viability of A549 and HCC827 cells was
determined by Cell Counting Kit-8 assay. (D) Analysis of ROS level
in A549 cells. (E) The levels of MDA, GSH and the ratio of GSH/GSSG
in A549 cells were detected by ELISA. *P<0.05 vs. control group;
#P<0.05 vs. TQ/HP-β-CD group. TQ, thymoquinone;
HP-β-CD, hydroxypropyl-β-cyclodextrin; ROS, reactive oxygen
species; MDA, malondialdehyde; GSH, reduced glutathione; GSSG,
oxidized glutathione.

Figure 9

TQ/HP-β-CD triggers ferroptosis in
the xenograft model of non-small-cell lung cancer. After treatment
with TQ and TQ/HP-β-CD in subcutaneous transplanted tumors, (A) the
growth curve, (B) tumor weight and (C) nude mouse weight were
measured. (D) ELISA method was used to detect the levels of MDA and
GSH in subcutaneous transplanted tumors. (E and F) Representative
images of H&E staining in liver, kidney, and tumor tissues were
used to evaluate the expression levels of Ki67 and NF-κB through
immunohistochemical staining. Prussian blue staining was used to
detect free iron deposition in tissues. *P<0.05 vs. control
group; #P<0.05 vs. TQ group. TQ, thymoquinone;
HP-β-CD, hydroxypropyl-β-cyclodextrin.
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Copy and paste a formatted citation
Spandidos Publications style
Zheng W, Ding J, Liang Y and Ye L: Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis. Oncol Rep 54: 157, 2025.
APA
Zheng, W., Ding, J., Liang, Y., & Ye, L. (2025). Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis. Oncology Reports, 54, 157. https://doi.org/10.3892/or.2025.8990
MLA
Zheng, W., Ding, J., Liang, Y., Ye, L."Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis". Oncology Reports 54.6 (2025): 157.
Chicago
Zheng, W., Ding, J., Liang, Y., Ye, L."Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis". Oncology Reports 54, no. 6 (2025): 157. https://doi.org/10.3892/or.2025.8990
Copy and paste a formatted citation
x
Spandidos Publications style
Zheng W, Ding J, Liang Y and Ye L: Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis. Oncol Rep 54: 157, 2025.
APA
Zheng, W., Ding, J., Liang, Y., & Ye, L. (2025). Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis. Oncology Reports, 54, 157. https://doi.org/10.3892/or.2025.8990
MLA
Zheng, W., Ding, J., Liang, Y., Ye, L."Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis". Oncology Reports 54.6 (2025): 157.
Chicago
Zheng, W., Ding, J., Liang, Y., Ye, L."Hydroxypropyl‑&beta;‑cyclodextrin/thymoquinone inclusion complex inhibits non‑small cell lung cancer progression through NF‑&kappa;B‑mediated ferroptosis". Oncology Reports 54, no. 6 (2025): 157. https://doi.org/10.3892/or.2025.8990
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