Department of Oral Pathology and Oral Radiology, Institute of Dentistry, Faculty of Medicine, University of Turku, Lemminkäisenkatu 2, FIN-20520, Turku, Finland.
Department of Pathology, Turku University Hospital, Turku, Finland.
Virol J. 2020 Jun 30;17(1):87. doi: 10.1186/s12985-020-01367-1.
This study was designed to investigate the invasion of human papillomavirus (HPV) positive human cervical carcinoma cell lines in human leiomyoma-based extracellular matrices in vitro, and to test the suitability of the model for studying the irradiation effects on the cancer cell invasion.
HPV positive cervical carcinoma cell lines SiHa and CaSki, and HPV negative squamous cell carcinoma cell line HSC-3 were used. CaSki cells contain around 600 copies of HPV 16 virus in the genome, whereas SiHa have only 1-2 copies per cell. Cells were analyzed using two different human tumor derived extracellular matrix methods (3D myoma disc model, and Myogel Transwell invasion assay). Cultures were irradiated with 4 Gy. Myoma invasion area and the depth of invasion were measured with ImageJ 1.51j8 software. Statistical analyses were performed with SPSS Statistics (IBM SPSS® Statistics 25).
All cells invaded through Myogel coated Transwell membranes and within myoma discs. In myoma discs, a difference in the invasion depth (p = 0.0001) but not in invasion area (p = 0.310) between the HPV positive cell lines was seen, since SiHa (less HPV) invaded slightly better than CaSki (more HPV). HSC-3 cells (HPV negative) invaded deepest (p = 0.048) than either of the HPV positive cell line cells. No difference was detected in the invasion area (p = 0.892) between HPV positive and HPV negative cells. The ionized radiation significantly reduced the invasion depth of HSC-3 (p = 0.008), SiHa (p = 0.0001) and CaSki (p = 0.005). No significant effect on the invasion area was detected in any of the cell lines. However, a significant difference was observed between SiHa and CaSki in the reduction of the invasion depth after radiation (p = 0.013) as the reduction was greater with SiHa than CaSki.
Both solid and gelatinous human leiomyoma-based extracellular matrix models were suitable platforms to study the invasion of HPV positive cervical carcinoma cells in vitro. SiHa cells with less HPV copy number cells invaded slightly better and were slightly more sensitive to irradiation than CaSki cells with high HPV copy number. However, there was no drastic differences between the invasion properties of these carcinoma cells.
本研究旨在体外研究人乳头瘤病毒（HPV）阳性人宫颈癌细胞系在人子宫肌瘤细胞外基质中的侵袭，并检验该模型用于研究辐射对癌细胞侵袭影响的适用性。
本研究使用了 HPV 阳性宫颈癌细胞系 SiHa 和 CaSki，以及 HPV 阴性鳞状细胞癌细胞系 HSC-3。CaSki 细胞的基因组中约有 600 个 HPV 16 病毒拷贝，而 SiHa 细胞每个细胞只有 1-2 个拷贝。采用两种不同的人肿瘤衍生细胞外基质方法（3D 肌瘤盘模型和 Myogel Transwell 侵袭测定）对细胞进行分析。细胞用 4Gy 进行照射。使用 ImageJ 1.51j8 软件测量 Myogel 包被的 Transwell 膜和肌瘤盘中的肿瘤侵袭面积和侵袭深度。采用 SPSS Statistics（IBM SPSS® Statistics 25）进行统计学分析。
所有细胞均穿过 Myogel 包被的 Transwell 膜并侵袭至肌瘤盘中。在肌瘤盘中，HPV 阳性细胞系之间的侵袭深度有差异（p=0.0001），但侵袭面积无差异（p=0.310），因为 SiHa（HPV 拷贝较少）的侵袭略优于 CaSki（HPV 拷贝较多）。HSC-3 细胞（HPV 阴性）的侵袭深度最深（p=0.048），明显大于任何 HPV 阳性细胞系细胞。HPV 阳性和 HPV 阴性细胞之间的侵袭面积无差异（p=0.892）。电离辐射显著降低了 HSC-3（p=0.008）、SiHa（p=0.0001）和 CaSki（p=0.005）的侵袭深度。然而，任何细胞系的侵袭面积均未检测到显著影响。但是，在辐射后 SiHa 和 CaSki 侵袭深度的降低之间观察到显著差异（p=0.013），因为 SiHa 的降低程度大于 CaSki。
固体和胶状人子宫肌瘤衍生细胞外基质模型均适合用于体外研究 HPV 阳性宫颈癌细胞的侵袭。HPV 拷贝数较少的 SiHa 细胞侵袭略好，对辐射的敏感性略高于 HPV 拷贝数较高的 CaSki 细胞。然而，这些癌细胞的侵袭特性之间没有明显差异。
