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背景与目的:HIF-1α是一个在缺氧状态下重要的转录调节因子,控制调节各种由缺氧引起的相关基因表达,参与肿瘤的恶化、浸润和转移,在肿瘤领域已成为研究热点。本文主要研究CoCl2模拟化学缺氧复氧中HIF-1α的表达,及其对人卵巢癌HO-8910PM细胞株生物学行为的影响。方法:在培养基中加入和去除CoCl2模拟化学缺氧和缺氧复氧,采用逆转录-聚合酶链反应(RT-PCR)检测CoCl2与HIF-1α的mRNA转录的量-效和时-效关系;通过MTT、Boyden小室、细胞黏附试验分别检测细胞生长、侵袭力和黏附力。结果:人卵巢癌HO-8910PM细胞株中存在HIF-1α的mRNA转录;在CoCl2模拟缺氧的时-效关系实验中,150μmol/LCoCl2刺激细胞8h、16h、24h时HIF-1αmRNA转录呈递增(分别为2.11±0.03、2.52±0.05、2.78±0.11)(P<0.05),24h达高峰,48h明显下降。同时,在量-效关系实验中,100μmol/L、150μmol/L刺激细胞16h时HIF-1αmRNA转录呈递增(分别为1.54±0.03、2.07±0.13)(P<0.05);MTT发现CoCl2诱导的缺氧对细胞生长起抑制作用;Boyden小室检测细胞侵袭力实验发现缺氧16h复氧24h后细胞的侵袭力增加(穿越细胞膜的细胞数86±9,较对照组53±10增加)(P<0.05);细胞黏附实验发现缺氧16h复氧24h后细胞的黏附力增加(P>0.05)。结论:CoCl2能诱导HO8910-PM细胞HIF-1α的过度转录,而且存在一定的时-效关系;经缺氧后复氧,肿瘤细胞的侵袭力和黏附力增加,且肿瘤细胞的侵袭力增加统计学差异显著。
BACKGROUND & OBJECTIVE: HIF-1α is an important transcriptional regulator in hypoxia. It regulates the expression of related genes induced by hypoxia and participates in the progression, invasion and metastasis of tumor. HIF-1α has become a hot topic in the field of oncology. In this paper, we mainly study the expression of HIF-1αin CoCl2 simulated hypoxia-reoxygenation and its effect on the biological behavior of human ovarian cancer cell line HO-8910PM. Methods: CoCl2 was used to simulate chemical hypoxia and hypoxia-reoxygenation in culture medium, and the amount-effect and time-effect of CoCl2 and HIF-1α mRNA transcription were detected by reverse transcription-polymerase chain reaction The cell growth, invasiveness and adhesion were detected by MTT, Boyden chamber and cell adhesion assay respectively. Results: HIF-1αmRNA transcription existed in human ovarian cancer HO-8910PM cell line. In the time-effect relationship of CoCl2 simulated hypoxia, HIF-1αmRNA transcription was increased at 150μmol / L NaCl for 8h, 16h and 24h Respectively, 2.11 ± 0.03,2.52 ± 0.05,2.78 ± 0.11) (P <0.05), reached the peak at 24 hours and decreased significantly at 48 hours. At the same time, HIF-1αmRNA transcripts were increased (P <0.05) at 100μmol / L and 150μmol / L for 16h (P <0.05) Oxygen inhibited the cell growth. Boyden’s cell invasion assay showed that invasiveness of cells was increased after 24 h of hypoxia (24h), the number of cells crossing the cell membrane was 86 ± 9 (53 ± 10 vs. control group) (P <0.05 ); Cell adhesion experiments found that after 24h hypoxia reoxygenation cell adhesion increased (P> 0.05). CONCLUSION: CoCl2 can induce the over-transcription of HIF-1α in HO8910-PM cells, and there is a certain time-effect relationship. After hypoxia and reoxygenation, the invasion and adhesion of tumor cells increase, and the invasiveness of tumor cells increases The difference is significant.