| Objective:This paper is aim to explore the influence of MHT on uterine leiomyomas in perimenopausal and postmenopausal women through an experimental Study. The study includes two parts:the influence of different types of MHT on uterine leiomyoma cells in same size and the influence of a certain MHT on uterine leiomyoma cells in different sizes. Furthermore, providing the Recommendations on MHT with experimental proof when it comes to "window period" women with uterine leiomyomas.Methods:(1) Uterine fibroid tissues were obtained from patients who underwent hysterectomy for multiple uterine leiomyoms (<3cm,3-5cm,>5cm included) in Zhongda Hospital from September 2015 to March 2016. The diagnosis of uterine leiomyomas was pathologically demonstrated. These perimenopausal and postmenopausal women (ranging from 45 to 55) who are younger than 60 years, within 10 years of menopause don’t receive MHT beceuse of suffering from uterine leiomyomas. Informed consent was obtained from each patient before surgery with the use of consent forms and protocols approved by the Ethics Committee of the Zhongda hospital. Choose three types of uterine leiomyoma tissues after pathological diagnosis, one cubic centimeter in size,5 samples each groups (Group A,B,C). Culture uterine leiomyoma tissues with collagenase digestion and build cell lines.(2) Drug intervention is done to the uterine leiomyoma cell lines. According to the types of MHT, we divide the uterine leiomyoma cells into three groups. A blank control group is set up as well. All the drugs are prepared to the concentrations below:1) Estradiol Valerate 30pg/ml; 2) Dydrogesterone 10ng/ml; 3) Estradiol Valerate full cycle+Dydrogesterone the second half of the cycle 30pg/ml+10ng/ml. The medium of four groups used is DMEM with 10% FBS. After 48h, the proliferation of cultured human uterine fibroid cells was analyzed by MTT assay and flow cytometry. Then explore the influence of different types of MHT on uterine leiomyoma cells in same size and the influence of a certain MHT on uterine leiomyoma cells in different sizes.Results:(1) Limited human uterine leiomyoma cells were successfully established and were identified by immunohistochemical method.(2) Human uterine leiomyoma cells were intervened by three kinds of drugs and culture medium for 48 hours,then the proliferation was analyzed by MTT assay. When the samples come from the same size of uterine leiomyomas, Estradiol Valerate 30pg/ml, Dydrogesterone 10ng/ml, Estradiol Valerate 30pg/ml full cycle plus Dydrogesterone 10ng/ml the second half of the cycle all can promote proliferation of uterine leiomyoma cells compared to the control group,the difference has statistics meaning (p<0.05); Estradiol Valerate, Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting the proliferation of uterine leiomyoma cells than Dydrogesterone, the difference has statistics meaning (p<0.05); Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting the proliferation of uterine leiomyoma cells than Estradiol Valerate, the difference has statistics meaning (p<0.05).(3) When intervened by the same drug pattern, the proliferation of uterine leiomyoma cells which comes from three different types(<3cm,3-5cm,>5cm) of uterine leiomyomas had no significant statistical differences (p>0.05).(4) Estradiol Valerate 30pg/ml, Dydrogesterone 10ng/ml, Estradiol Valerate 30pg/ml full cycle plus Dydrogesterone 10ng/ml the second half of the cycle all can promote uterine leiomyoma cells into S cell cycle phase and elevated mitotic index compared to the control group, the difference has statistics meaning (p<0.05); Estradiol Valerate, Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting uterine leiomyoma cells into S cell cycle phase and elevated mitotic index than Dydrogesterone, the difference has statistics meaning (p<0.05); Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting uterine leiomyoma cells into S cell cycle phase and elevated mitotic index than Estradiol Valerate, the difference has statistics meaning (p<0.05).Conclusions:(1)When the samples come from the same size of uterine leiomyoma, Estradiol Valerate, Dydrogesterone, Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle all can promote proliferation of uterine leiomyoma cells.Estradiol Valerate, Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting the proliferation of uterine leiomyoma cells than Dydrogesterone. Estradiol Valerate full cycle plus Dydrogesterone the second half of the cycle was more effective in promoting the proliferation of uterine leiomyoma cells than Estradiol Valerate.(2) When intervened by the same drug patter, the proliferation of uterine leiomyoma cells which corned from three different tyes of uterine leiomyomas had no significant statistical differences.(3) The drugs of MHT can affect the cell cycles of cultured uterine leiomyoma cells and promote the proliferation of the cells. |
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