J Gynecol Oncol.  2011 Dec;22(4):225-232. 10.3802/jgo.2011.22.4.225.

Combination of imatinib mesylate with lithium chloride and medroxyprogesterone acetate is highly active in Ishikawa endometrial carcinoma in vitro

• Bilir A
• Erguven M
• Ermis E
• Sencan M
• Yazihan N

• KMID: 2173623
• DOI: http://doi.org/10.3802/jgo.2011.22.4.225

Abstract

OBJECTIVE
The aim of the study was to investigate whether lithium chloride and medroxyprogesterone acetate can potentiate the cytotoxicity of imatinib mesylate in human endometrial cancer in vitro and the effect of midkine in these therapies.
METHODS
Imatinib mesylate (50 microM), lithium chloride (100 microM), medroxyprogesterone acetate (200 microM) and their combination were applied to monolayer and three dimensional cultures of human Ishikawa endometrial cancer for 72 hours. The cell proliferation index, apoptotic index, caspase-3 and midkine levels, cell cycle distributions in monolayer cultures and cell ultrastructure in spheroid cultures were evaluated. Results were statistically analyzed using the Student's t-test.
RESULTS
All drug applications inhibited cell proliferation (p<0.05), however the combination were the effective groups for 72 hours (p<0.05). Interestingly, although the loss of efficiency was seen higly seen every 24 hours at single applications, the inhibition rates of the combination groups were almost same for 72 hours. In concordance with these results, the apoptotic index, caspase-3 levels (p<0.05), cell morphology and ultrastructure damages were much higher in the combination groups. Imatinib mesylate induced S-phase arrest, however other groups induced G0+G1-phase arrest at 24 hours and all groups induced G0+G1 arrest at 72 hours (p<0.05). Imatinib mesylate and imatinib mesylate with medroxyprogesterone acetate induced highest decrease in midkine levels, respectively (p<0.05).
CONCLUSION
The present study showed that the combination of imatinib mesylate with lithium chloride and medroxyprogesterone acetate is highly active in Ishikawa endometrial carcinoma in vitro and the inhibition of midkine involved in their mechanism of action against endometrium defense.

Keyword

Endometrial cancer; Imatinib mesylate; Lithium chloride; Medroxyprogesterone acetate; Midkine

MeSH Terms

Benzamides
Caspase 3
Cell Cycle
Cell Proliferation
Cytokines
Endometrial Neoplasms
Endometrium
Female
Humans
Imatinib Mesylate
Lithium
Lithium Chloride
Medroxyprogesterone
Medroxyprogesterone Acetate
Mesylates
Piperazines
Pyrimidines
Benzamides
Caspase 3
Cytokines
Lithium
Lithium Chloride
Medroxyprogesterone
Medroxyprogesterone Acetate
Mesylates
Piperazines
Pyrimidines

Figure

• 

  Fig. 1 The effects of single and combined drugs on cell proliferation. Columns, average of six wells. Data are representative of separate three experiments. C, control; IM, imatinib mesylate; LiCl, lithium chloride; IM+LiCl, the combination of IM and LiCl; MPA, medroxyprogesterone acetate; IM+MPA, the combination of IM and MPA.

• 

  Fig. 2 The alterations in apoptotic index induced by different drug applications. Columns, average of six wells. Data are representative of separate three experiments. C, control; IM, imatinib mesylate; LiCl, lithium chloride; IM+LiCl, the combination of IM and LiCl; MPA, medroxyprogesterone acetate; IM+MPA, the combination of IM and MPA.

• 

  Fig. 3 Caspase-3 levels effected by single and combined drugs. Columns, average of six wells. Data are representative of separate three experiments. C, control; IM, imatinib mesylate; LiCl, lithium chloride; IM+LiCl, the combination of IM and LiCl; MPA, medroxyprogesterone acetate; IM+MPA, the combination of IM and MPA.

• 

  Fig. 4 The alterations of cell cycle distributions by different drug applications. Columns, average of six wells. Data are representative of separate three experiments. C, control; IM, imatinib mesylate; LiCl, lithium chloride; IM+LiCl, the combination of IM and LiCl; MPA, medroxyprogesterone acetate; IM+MPA, the combination of IM and MPA.

• 

  Fig. 5 The effect of different drug applications on MDK levels. Columns, average of six wells. Data are representative of separate three experiments. C, control; IM, imatinib mesylate; LiCl, lithium chloride; IM+LiCl, the combination of IM and LiCl; MPA, medroxyprogesterone acetate; IM+MPA, the combination of IM and MPA.

• 

  Fig. 6 The alterations in cell ultrastructure by different drug applications. (A) Control (×7,500), (B) Imatinib mesylate (IM; ×10 k), (C) Lithium chloride (LiCl; ×6,000 and ×6,000), (D) IM+LiCl (×10 k and ×7,500), (E) Medroxyprogesterone acetate (MPA; ×7,500), (F) IM+MPA (×10 k and ×7,500). n, nucleus; nu, nucleolus; lp, lipid vacuole; mi, mitochondria; mv, microvillus; lt, lytic cytoplasm; j, junctions; v, vacuole; ch, chromatin; cr, cell remnants; *, apoptotic cell; av, autophagic vacuole; g, gaps.

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