褪黑素对培养的人卵巢癌细胞增殖及顺铂（CDDP）敏感性的影响。

Effects of melatonin on the proliferation and cis-diamminedichloroplatinum (CDDP) sensitivity of cultured human ovarian cancer cells.

作者信息

Futagami M, Sato S, Sakamoto T, Yokoyama Y, Saito Y

机构信息

Department of Obstetrics and Gynecology, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036-8562, Japan.

出版信息

Gynecol Oncol. 2001 Sep;82(3):544-9. doi: 10.1006/gyno.2001.6330.

DOI:10.1006/gyno.2001.6330

PMID:11520153

Abstract

OBJECTIVES

In this study, we investigated the antiproliferative effect and telomerase activity of melatonin with or without cis-diamminedichloroplatinum (CDDP) on CDDP-sensitive HTOA cells and CDDP-resistant OVCAR-3 cells of cultured human ovarian cancer.

METHODS

HTOA cells and OVCAR-3 cells were cultured in RPMI-1640 at 37 degrees C with 5% CO(2) for 132 h. To examine the antiproliferative effect of melatonin, cells were cultured with or without melatonin (10(-12)-10(-6) M) and thereafter counted in a 96-well microplate using the alamarBlue assay. To examine the effect of melatonin and CDDP, cells were divided into group A (intermittent CDDP, 0.5 microg/ml), group B (intermittent CDDP + melatonin), and group C (sequential (12-h interval) CDDP/melatonin) and thereafter counted in a 96-well microplate using the alamarBlue assay. In different series, cells were cultured and treated with either ethanol, melatonin, CDDP, or CDDP + melatonin. After harvest, telomerase activity was semiquantified with a fluorescence-based telomeric repeat amplification protocol (F-TRAP).

RESULTS

(1) Melatonin produced no antiproliferative effect on both types of ovarian cancer cells. (2) Melatonin 10(-6) M induced the antiproliferative effect in groups B and C compared with group A in the HTOA cell line. (3) Melatonin 10(-9) M produced the antiproliferative effect in groups B and C compared with group A in the OVCAR-3 cell line. (4) Telomerase activity in the HTOA cell line did not change but, in the OVCAR-3 cell line, was significantly lower in the CDDP + melatonin group compared with the ethanol and CDDP groups.

CONCLUSIONS

Melatonin enhanced CDDP sensitivity in two ovarian cancer cell lines. Thus, melatonin may improve ovarian cancer chemotherapy.

摘要

目的

在本研究中，我们调查了褪黑素单独或联合顺二氨二氯铂（CDDP）对培养的人卵巢癌CDDP敏感的HTOA细胞和CDDP耐药的OVCAR - 3细胞的抗增殖作用及端粒酶活性。

方法

将HTOA细胞和OVCAR - 3细胞在含5% CO₂的RPMI - 1640培养基中于37℃培养132小时。为检测褪黑素的抗增殖作用，细胞分别在有或无褪黑素（10⁻¹² - 10⁻⁶ M）的条件下培养，之后使用alamarBlue检测法在96孔微孔板中进行细胞计数。为检测褪黑素和CDDP的作用，细胞被分为A组（间歇给予CDDP，0.5μg/ml）、B组（间歇给予CDDP + 褪黑素）和C组（序贯给予（间隔12小时）CDDP/褪黑素），之后使用alamarBlue检测法在96孔微孔板中进行细胞计数。在不同系列实验中，细胞分别用乙醇、褪黑素、CDDP或CDDP + 褪黑素进行培养和处理。收获细胞后，采用基于荧光的端粒重复序列扩增法（F - TRAP）对端粒酶活性进行半定量分析。

结果

（1）褪黑素对两种类型的卵巢癌细胞均无抗增殖作用。（2）在HTOA细胞系中，与A组相比，10⁻⁶ M的褪黑素在B组和C组诱导产生了抗增殖作用。（3）在OVCAR - 3细胞系中，与A组相比，10⁻⁹ M的褪黑素在B组和C组产生了抗增殖作用。（4）HTOA细胞系中的端粒酶活性未发生变化，但在OVCAR - 3细胞系中，与乙醇组和CDDP组相比，CDDP + 褪黑素组的端粒酶活性显著降低。