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PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

  • Authors:
    • Haiyan Wang
    • Ruiming Zheng
    • Bujun Wang
    • Sumiao Hong
    • Hongxia Chen
  • View Affiliations / Copyright

    Affiliations: Department of Obstetrics and Gynecology, Pingyang People's Hospital, Wenzhou, Zhejiang 325400, P.R. China
    Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 278
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    Published online on: August 14, 2026
       https://doi.org/10.3892/etm.2026.13273
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Abstract

Ovarian cancer (OC) remains one of the most lethal gynecologic malignancies, with limited therapeutic options and poor long‑term prognosis. Gramine, an indole alkaloid predominantly derived from the rhizomes of Arundo donax L., has demonstrated antitumor activity in various cancer types; however, its effects and mechanistic basis in OC have not been systematically investigated. In the present study, the biological activity of gramine in OC cells were evaluated using cell counting kit‑8, colony formation and EdU assays for proliferation; TUNEL staining for apoptosis; and wound healing and Transwell assays for migration and invasion. Western blotting was performed to examine underlying molecular changes. The present results showed that gramine significantly suppressed OC cell proliferation, migration and invasion, while promoting cell apoptosis. Mechanistically, gramine reduced PDK1 protein levels, and enforced PDK1 overexpression largely reversed the suppressive effects of gramine on malignant phenotypes. Consistently, PDK1 restoration also attenuated gramine‑induced downregulation of phosphorylated AKT. Furthermore, gramine enhanced the sensitivity of OC cells to cisplatin, an effect associated with PDK1/AKT signaling inhibition. Collectively, these findings suggest that gramine inhibits malignant progression and attenuates cisplatin resistance in OC cells, at least in part, through blockade of the PDK1/AKT pathway. Although limited to in vitro models, this work provides preliminary evidence supporting further exploration of gramine as a potential research tool or lead compound in OC.
View Figures

Figure 1

Gramine suppressed cell proliferation
in ovarian cancer cells. (A) The chemical structure of gramine. (B)
Cell counting kit-8 assay detecting the effect of gramine on cell
viability. SK-OV-3 and OV-90 cells were incubated with gramine
(12.5, 25, 50, 100, 200 µM) for 24 h. (C) Clone formation assay
assessing the effect of gramine on clone formation ability. SK-OV-3
and OV-90 cells were incubated with gramine (50, 100 µM) for 72 h.
(D) EdU assay measuring the effect of gramine on the cell
proliferation. Scale bar, 200 µm. (E) Western blotting assay
testing the effect of gramine on Ki67 expressions. SK-OV-3 and
OV-90 cells were incubated with gramine (50, 100 µM) for 24 h.
**P<0.01, ***P<0.001 vs. the untreated
control. Data represent the mean ± SD of three independent
experiments.

Figure 2

Gramine induced apoptosis in OC cells.
(A) TUNEL assay evaluating the effect of gramine on cell apoptosis.
Scale bar, 200 µm. (B) Western blotting assay determining the
effect of gramine on Bax, Bcl2, PARP1 and cleaved PARP1
expressions. SK-OV-3 and OV-90 cells were incubated with gramine
(50,100 µM) for 24 h. *P<0.05,
**P<0.01, ***P<0.001 vs. the untreated
control. Data represent the mean ± SD of three independent
experiments.

Figure 3

Gramine inhibited cell migration and
invasion in OC cells. (A) Wound healing and (B) transwell invasion
assays detecting the effect of gramine on the abilities of cell
migration and invasion respectively. Scale bar, 200 µm. SK-OV-3 and
OV-90 cells were incubated with gramine (50 µM) for 24 h. (C)
Western blotting assay determining the effect of gramine on
N-cadherin and vimentin expressions. SK-OV-3 and OV-90 cells were
incubated with gramine (50, 100 µM) for 24 h.
*P<0.05, **P<0.01,
***P<0.001 vs. the untreated control. Data represent
the mean ± SD of three independent experiments.

Figure 4

PDK1 overexpression impaired the
anti-proliferation effect of gramine on OC cells. (A) Western
blotting assay determining the effect of gramine on PDK1
expressions. SK-OV-3 and OV-90 cells were incubated with gramine
(50, 100 µM) for 24 h. *P<0.05,
**P<0.01, ***P<0.001 vs. the untreated
control. (B) Western blotting assay assessing the transfection
efficiency of PDK1 overexpression lentivirus.
***P<0.001 vs. the EV control. (C) Cell counting
kit-8 assay detecting the effect of PDK1 overexpression on cell
viability. (D) Clone formation assay assessing the effect of PDK1
overexpression on colony formation ability. (E) Western blotting
assay examining the effect of PDK1 overexpression on Ki67
expression. *P<0.05, **P<0.01 vs. the
EV control or gramine-treated EV group (C-E). Data represent the
mean ± SD of three independent experiments. EV, empty vector; OV,
overexpression.

Figure 5

PDK1 overexpression impaired the
pro-apoptotic effect of gramine on ovarian cancer cells. (A) TUNEL
assay evaluating the effect of PDK1 overexpression on cell
apoptosis. Scale bar, 200 µm. (B) Western blotting assay
determining the effect of PDK1 overexpression on Bax and Bcl2
expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were
treated with gramine (100 µM) for 24 h. *P<0.05,
**P<0.01, ***P<0.001 vs. the EV control
or gramine-treated EV group. Data represent the mean ± SD of three
independent experiments. EV, empty vector; OV, overexpression.

Figure 6

PDK1/AKT signaling pathway was
involved in the anti-migratory and anti-invasive effects of gramine
in ovarian cancer cells. (A) Wound healing and (B) transwell
invasion assays detecting the effect of PDK1 overexpression on the
abilities of cell migration and invasion respectively. Scale bar,
200 µm. (C) Western blotting assay determining the effect of PDK1
overexpression on N-cadherin, vimentin and p-AKT (Thr308)
expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were
incubated with gramine (50 µM) for 24 h. *P<0.05,
**P<0.01, ***P<0.001 vs. the EV control
or gramine-treated EV group. Data represent the mean ± SD of three
independent experiments. EV, empty vector; OV, overexpression.

Figure 7

Gramine downregulates PDK1/AKT linked
to cisplatin sensitization of OC cells. (A) CCK-8 assay detecting
the effect of cisplatin on cell viability. SK-OV-3 and SK-OV-3-R
cells were incubated with cisplatin (5, 10, 20, 40, 80 µM) for 24
h. *P<0.05, **P<0.01 vs. the untreated
control. (B) CCK-8 assay detecting the effect of gramine and
cisplatin on cell viability. SK-OV-3-R cells were incubated with
gramine (50 µM) and cisplatin (5, 10, 20, 40, 80 µM) for 24 h.
*P<0.05 vs. the DMSO group at the identical cisplatin
concentration. (C) Clone formation assay assessing the effect of
gramine and cisplatin on colony formation ability. (D) TUNEL assay
evaluating the effect of gramine and cisplatin on cell apoptosis.
Scale bar, 200 µm. (E) Wound healing and transwell invasion assays
detecting the effect of gramine and cisplatin on the abilities of
cell migration and invasion respectively. Scale bar, 200 µm. (F)
Western blotting assay determining the effect of gramine and
cisplatin on the expressions of PDK1 and p-AKT (Thr308). SK-OV-3-R
cells were incubated with gramine (50 µM) and cisplatin (80 µM) for
24 h. *P<0.05, **P<0.01,
***P<0.001 vs. the cisplatin group (C-F). Data
represent the mean ± SD of three independent experiments. CCK-8,
cell counting kit-8.
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Copy and paste a formatted citation
Spandidos Publications style
Wang H, Zheng R, Wang B, Hong S and Chen H: PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells. Exp Ther Med 32: 278, 2026.
APA
Wang, H., Zheng, R., Wang, B., Hong, S., & Chen, H. (2026). PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells. Experimental and Therapeutic Medicine, 32, 278. https://doi.org/10.3892/etm.2026.13273
MLA
Wang, H., Zheng, R., Wang, B., Hong, S., Chen, H."PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells". Experimental and Therapeutic Medicine 32.4 (2026): 278.
Chicago
Wang, H., Zheng, R., Wang, B., Hong, S., Chen, H."PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells". Experimental and Therapeutic Medicine 32, no. 4 (2026): 278. https://doi.org/10.3892/etm.2026.13273
Copy and paste a formatted citation
x
Spandidos Publications style
Wang H, Zheng R, Wang B, Hong S and Chen H: PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells. Exp Ther Med 32: 278, 2026.
APA
Wang, H., Zheng, R., Wang, B., Hong, S., & Chen, H. (2026). PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells. Experimental and Therapeutic Medicine, 32, 278. https://doi.org/10.3892/etm.2026.13273
MLA
Wang, H., Zheng, R., Wang, B., Hong, S., Chen, H."PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells". Experimental and Therapeutic Medicine 32.4 (2026): 278.
Chicago
Wang, H., Zheng, R., Wang, B., Hong, S., Chen, H."PDK1/AKT signaling is involved in the anti‑tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells". Experimental and Therapeutic Medicine 32, no. 4 (2026): 278. https://doi.org/10.3892/etm.2026.13273
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