R-spondin3 promotes the tumor growth of choriocarcinoma JEG-3 cells

Abstract

R-spondin3 (RSPO3), an activator of Wnt/β-catenin signaling, plays a key role in tumorigenesis of various cancers, but its role in choriocarcinoma remains unknown. To investigate the effect of RSPO3 on the tumor growth of choriocarcinoma JEG-3 cells, the expression of RSPO3 in human term placenta was detected, and a stable RSPO3-overexpressing JEG-3 cell line was established via lentivirus-mediated transduction. The expression of biomarkers involved in tumorigenicity was detected in the RSPO3-overexpressing JEG-3 cells, and cell proliferation, invasion, migration, and apoptosis were investigated. Moreover, soft agar clonogenic assays and xenograft tumorigenicity assays were performed to assess the effect of RSPO3 on tumor growth in vitro and in vivo. The results showed that RSPO3 was widely expressed in human term placenta and overexpression of RSPO3 promoted the proliferation and inhibited the migration, invasion, and apoptosis of the JEG-3 cells. Meanwhile, RSPO3 overexpression promoted tumor growth both in vivo and in vitro. Further investigation showed that the phosphorylation levels of Akt, phosphatidylinositol 3-kinase (PI3K), and ERK as well the expression of β-catenin and proliferating cell nuclear antigen (PCNA) were increased in the RSPO3-overexpressing JEG-3 cells and tumor xenograft. Taken together, these data indicate that RSPO3 promotes the tumor growth of choriocarcinoma via Akt/PI3K/ERK signaling, which supports RSPO3 as an oncogenic driver promoting the progression of choriocarcinoma.

α-Solanine Inhibits Proliferation, Invasion, and Migration, and Induces Apoptosis in Human Choriocarcinoma JEG-3 Cells In Vitro and In Vivo

“…In the colony formation assay, the number of cells was small (only 200 cells per well), which may increase the sensitivity of cells to α-solanine. Moreover, an animal model was established using malignant trophoblast JEG-3 cells grown in nude mice, in which fast-growing tumors developed in a short time [25,26]. The growth of the choriocarcinoma xenografts was significantly inhibited by α-solanine.…”

“…Proteolytic enzymes, like MMPs, are closely associated with disease progression and metastasis due to the degradation of the ECM in many cancers [34,35]. The inhibition of MMP-2/9 reduces tumor invasion and metastasis in cervical cancer cells [36,37], pancreatic cancer [38], and choriocarcinoma [26]. The levels of MMP-2/9 mRNA, as well inactive MMP-2 (pro-MMP-2), were decreased in α-solanine-treated JEG-3 cells, indicating that the inhibitory effect of α-solanine on the metastasis of choriocarcinoma cells may be associated with MMP-2/9.…”

“…The culture of JEG-3 cells (a human choriocarcinoma cell line) has been described previously [26]. To study the cytotoxic effect of α-solanine on JEG-3 cells, cells were cultured in 96-well plates overnight, followed by treatment with different concentrations of αsolanine (10, 20, 30, 40 and 50 µM) or vehicle (0.5% v/v DMSO) for 24 h. Then, MTS reagent was added (20 µL each well) and incubated at 37 • C for 3 h. The absorbance at 490 nm was measured using a microplate reader (NanoQuant Infinite M200 Pro, Tecan, Mannedorf, Switzerland).…”

“…A wound healing assay was performed to assess cellular migratory ability as described previously [26]. Briefly, JEG-3 cells were seeded in 6-well plates.…”

“…A Matrigel invasion assay was performed to evaluate the invasive ability of JEG-3 cells as described previously with minor modifications [26]. JEG-3 cells in serum-free medium with 30 µM α-solanine or vehicle were seeded in the upper surface of Matrigelcoated Transwell inserts.…”