Anat Cell Biol.  2021 Mar;54(1):104-111. 10.5115/acb.20.228.

Anti-proliferative and apoptotic effects of hull-less pumpkin extract on human papillary thyroid carcinoma cell line

• Bahadori MH
• Azari Z
• Zaminy A
• Dabirian S
• Mehrdad SM
• Kondori BJ

• KMID: 2514590
• DOI: http://doi.org/10.5115/acb.20.228

Abstract

Papillary thyroid carcinoma (PTC) is one of the most common cancers of the endocrine system. Previous studies have shown that the extract of hull-less pumpkin seed (HLPS) has a significant anti-cancer effect. The aim of this study was to evaluate the effect of this plant extract on the proliferation of PTC cells. In this study, an extract of this plant was prepared by soxhlet extraction method and analyzed by Gas Chromatography-Mass Spectrometry. The cytotoxicity of PTX and plant extract was investigated using the methylthiazol tetrazolium (MTT) method. For careful investigation of morphological alteration, we used hematoxylin and eosin and Giemsa stinging. Based on MTT assay test, the IC 50 value of paclitaxel (PTX) was significantly less than the hydro-alcoholic extract of HLPS at all of the incubation time. Our results of histological staining showed that HLPS and PTX induced significant morphological alteration in the PTC cultured cell that consistent with cell death. Comparing the groups treated by PTX or HLPS with control group showed significant differences. It seems that HLPS extract has an apparent effect on treatment of PTC, at least in laboratory condition, albeit for realistic decision about the effect of HLPS on PTC, more molecular investigations are necessary.

Keyword

Thyroid neoplasms; Paclitaxel; Cell proliferation; Herbal medicine

Figure

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  Fig. 1 Chromatogram of hull less seed pumpkin extract. Most common compounds in the extract is 9,12-octadecadienoic acid ethyl esters (Oleic Acid): 16.21%.

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  Fig. 2 The MTT assay results of treated cells by hydro-alcoholic extract of HLPS for 24 (A), 48 (B), and 72 (C) hours. In the dose of 1, 20, 50, 100, and 200 μg/ml the inhibition of PTC cell proliferation was 5% to 15% and in the dose of 800 μg/ml was nearly 30%. HLPS, hull-less pumpkin seed; MTT, methylthiazol tetrazolium; PTC, paclitaxel.

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  Fig. 3 The MTT assay results of PTX treated cells. The results show that this drug in the dose of 0.001, 0.0001, and 0.00001 μg/ml after 24 (A) and 48 (B) hours don’t inhibit cell proliferation. Also, in the dose of 0.01 and 0.1 μg/ml, PTX don’t effects on PTC cell proliferation significantly (P>0.05). in the dose of 0.1 μg/ml after 72 hours (C) the inhibition of cell proliferation was nearly 30%. MTT, methylthiazol tetrazolium; PTC, papillary thyroid carcinoma; PTX, paclitaxel.

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  Fig. 4 H&E staining of PTC cells. Control group without any treatment (A). PTC cells under treatment with HLPS with the dose of 800 μg/ml (B), and 1,600 μg/ml (C). PTC cells under treatment with PTX with the dose of 1 μg/ml (D); Chromatin condensation was occurred after cell treatment with different doses of PTX or HLPS hydro-alcoholic extract. The arrow heads indicate normal cells and the arrows indicate chromatin condensation and cell death. HLPS, hull-less pumpkin seed; PTC, papillary thyroid carcinoma; PTX, paclitaxel. Scale bar=10 μm.

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  Fig. 5 Giemsa staining; untreated PTC cells (A), PTC cells treated by HLPS extract (800 μg/ml) (B), and 1,600 μg/ml (C) after 24 hours. PTC cells treated with PTX (1 μg/ml), after 24 hourrs (D). Nuclear fragmentation and cell death were occurred after cell treatment with different doses of PTX or HLPS hydro-alcoholic extract (arrows). HLPS, hull-less pumpkin seed; PTC, papillary thyroid carcinoma; PTX, paclitaxel. Scale bar=10 μm.

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  Fig. 6 Compare the percentage of dead cells in PTC cells treated with extract and PTX at the 24, 48, and 72 hours. PTC, papillary thyroid carcinoma; PTX, paclitaxel.

Reference

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