miR-195-5p靶向FGF2 抑制子宫内膜癌HEC-1B细胞恶性生物学行为

目的：探讨miR-195-5p 通过靶向FGF2 抑制子宫内膜癌HEC-1B细胞增殖、凋亡、侵袭和迁移的分子机制。方法：HEC-1B细胞培养与转染完成后分为4 组：HEC-1B组、miR-195-5p mimic组、pLV-FGF2 组和miR-195-5p+FGF2 组。qRT-PCR检测细胞miR-195-5p 和FGF2 mRNA水平，荧光素酶实验验证miR-195-5p 与FGF2 的靶向关系，Western blotting 检测FGF2 表达水平，CCK-8 法检测HEC-1B细胞增殖水平，流式细胞术检测HEC-1B细胞凋亡率，Transwell 实验检测HEC-1B细胞侵袭能力，划痕实验检测HEC-1B细胞迁移能力。结果：与HEC-1B组相比，miR-195-5p mimic 组miR-195-5p 表达升高、FGF2 mRNA水平下降(均P< 0.01)；miR-195-5p 可直接靶向FGF2。与HEC-1B组相比，miR-195-5p mimic 组FGF2 的蛋白表达水平下降，pLV- FGF2 组FGF2 的蛋白水平明显上升，且miR-195-5p+FGF2 组FGF2 的蛋白表达水平低于pLV- FGF2 组(均P< 0.01)。miR-195-5p mimic 组细胞增殖水平低于HEC-1B组，pLV-FGF2 组细胞增殖水平高于HEC-1B组(均P< 0.01)。与HEC-1B组相比，miR-195-5p mimic 组细胞凋亡率增加，pLV- FGF2 组细胞凋亡率降低，且miR-195-5p+ FGF2 组细胞凋亡率高于pLV- FGF2 组(均P< 0.01)。与HEC-1B组相比，miR-195-5p mimic 组每个视野下的侵袭细胞数和划痕愈合率下降，pLV- FGF2 组每个视野下的侵袭细胞数和划痕愈合率上升，且miR-195-5p+FGF2 组每个视野下的侵袭细胞数和划痕愈合率低于pLV- FGF2 组(均P<0.01)。结论: miR-195-5p 通过靶向FGF2 抑制子宫内膜癌HEC-1B细胞的增殖、侵袭和迁移并促进细胞凋亡，其作为子宫内膜癌的治疗靶点。

MiR-195-5p targeting FGF2 inhibits malignant biological behaviors of endometrial carcinoma HEC-1B cells

Objective: To explore the molecular mechanism of miR-195-5p targeting FGF2 to inhibit the proliferation, apoptosis, invasion and migration of endometrial cancer HEC-1B cells. Methods: After culture and transfection, HEC-1B cells were divided into 4 groups: HEC-1B group, miR-195-5p mimic group, pLV-FGF2 group and miR-195-5p+FGF2 group. The expressions of miR-195-5p and mRNA levels of FGF2 were detected by qRT-PCR. The targeted relationship of miR-195-5p and FGF2 was verified by luciferase assay. The protein expression of FGF2 was examined by Western blotting; Proliferation of HEC-1B cells was measured by CCK-8;Apoptosis was tested by flow cytometry; HEC-1B cell invasion was detected by transwell, and migration was measured by scratch assay.Results: Compared with HEC-1B group, the expression of miR-195-5p in miR-195-5p mimic group was elevated while FGF2 mRNA level was declined (all P<0.01). Luciferase assay indicated that FGF2 was a target of miR-195-5p. Compared with HEC-1B group, the protein level of FGF2 in miR-195-5p mimic group was decreased, and the protein levels of FGF2 in pLV-FGF2 group were enhanced (P<0.01). The protein levels of FGF2 in miR-195-5p+FGF2 group were lower than that in pLV-FGF2 group (all P<0.01). The proliferation in miR-195-5p mimic group was lower than HEC-1B group (P<0.01), while the proliferation in pLV-FGF2 group was higher than that in HEC-1B group (all P<0.01). Compared with HEC-1B group, apoptosis in miR-195-5p mimic group was increased,and apoptosis in pLV-FGF2 group was decreased (P<0.01); moreover, apoptosis in miR-195-5p+FGF2 group was higher than that in pLV-FGF2 group (P<0.01). Compared with HEC-1B group, the number of invasive cells per field and the rate of wound healing in miR-195-5p mimic group were decreased, while those in pLV-FGF2 group was enhanced (P<0.01); moreover, the number of invasive cells per field and the rate of wound healing in miR-195-5p+FGF2 group was lower than those in pLV-FGF2 group (all P<0.01). Conclusion:miR-195-5p inhibits proliferation, invasion and migration and promotes apoptosis of endometrial cancer HEC-1B cells by target-ing FGF2, and could be used as a treatment target of endometrial cancer.