Study on mechanism of syncytiotrophoblast extracellular vesicles preventing maternal malignant tumor invasion and metastasis to fetus

doi:10.3877/cma.j.issn.1673-5250.2022.01.005

Objective

To explore the mechanism of syncytiotrophoblast extracellular vesicles (STB-EV) preventing maternal malignant tumor invasion and metastasis to fetus.

Methods

Placental tissue invaded by tumor of a pregnant woman with cervical cancer in pregnancy (CCP) who underwent cesarean section in West China Second University Hospital, Sichuan University in June 2015 was selected as research subject. Cervical cancer cell line SiHa cells and human trophoblast cell line HTR-8 cells were donated by the Laboratory of Molecular and Translational Medicine, Western Institute of Maternal and Child Medicine, West China Second University Hospital, Sichuan University. Placental tissue invaded by cervical cancer of this CCP patient was sectioned, and its histomorphology, STB-EV and autophagic death of cervical cancer cells adjacent to syncytiotrophoblast (STB), etc. were observed under optical microscope (×400) and electron microscope (×2 000). SiHa cells and HTR-8 cells were cultured for Transwell migration assay and cell scratch test. Co-cultured SiHa cells and HTR-8 cells were included into study group, while SiHa cells cultured separately were included into control group. Transwell migration assay and cell scratch test were performed on two groups of cells to evaluate the interaction between HTR-8 and SiHa cells. Independent-samples t test was used to compare the number of transmembrane penetrating cells and cell mobility between two groups. The procedures followed in this study were in line with the newly revised World Medical Association Declaration of Helsinki in 2013.

Results

①Results of placental tissue invaded by cervical cancer of this CCP patients under optical microscope (×400) showed that autophagy death, apoptosis and pyknotic necrosis of cervical cancer cells adjacent to STB were significant. Electron microscopy (×2 000) results showed that when cervical cancer cells involved placental villi, level of STB-EV secreted by STB was significantly higher than that in normal placental tissues. ②Results of Transwell migration assay in two groups showed that the number of transmembrane penetrating cells in study group was (597.6±87.7) field of view, which was significantly lower than that in control group (1 358.4±203.0) field of view, and the difference was statistically significant (t=14.490, P<0.001). ③Results of cell scratch test in two groups showed that cell migration rate at 48 h after scratch in study group was significantly lower than that in the control group [(26.6±3.8)% vs (45.9±3.7)%, t=3.122, P=0.035].

Conclusions

STB-EV secreted by STB may be involved in extensive autophagic death of maternal tumor cells, so as to prevent maternal tumor cells from invading placenta and affecting fetal development.

Key words: Trophoblasts, Syncytiotrophoblast extracellular vesicle, Cervix neoplasms, Placenta, Pregnancy complications, neoplastic, Pregnant women

图3 本例CCP患者(女性，33岁)宫颈癌组织切片于电子显微镜下观察结果，可见累及STB宫颈癌细胞较未累及STB宫颈癌细胞自噬小体增多(箭头所示)、坏死明显(图3A：累及STB的宫颈癌组织；图3B：未累及STB的宫颈癌组织)(高倍)

图4 光学显微镜下观察研究组与对照组中Transwell小室穿膜细胞数(图4A、4B：研究组不同区域穿膜细胞；图4C、4D：对照组不同区域穿膜细胞)(高倍)注：研究组为SiHa细胞与HTR-8细胞共培养细胞；对照组为单独培养的SiHa细胞

图5 研究组与对照组细胞划痕实验结果(图5A：研究组划痕时；图5B：对照组划痕时；图5C：研究组划痕48 h后；图5D：研究组划痕48 h后；图5E：研究组与对照组细胞迁移率比较)(高倍)注：研究组为SiHa细胞与HTR-8细胞共培养细胞；对照组为单独培养的SiHa细胞

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