A novel RGD-toxin protein, Lj-RGD3, from the buccal gland secretion of Lampetra japonica impacts diverse biological activities

Meriolin1 induces cell cycle arrest, apoptosis, autophagy and targeting the Akt/MAPKs pathways in human neuroblastoma SH-SY5Y cells

“…The extracellular matrix (ECM) protein matrigel (BD Biosciences) was used to coat the wells in the 96‐well plate for 20 min. After SH‐SY5Y cells were pretreated with Meriolin1 (0.5, 1, 2 μ m ) for 48 h, 1 × 10 6 cells/ml of cell suspension was seeded into the precoated 96‐well plate and incubated at 37 °C for 2 h. When the unbound SH‐SY5Y cells were washed twice with PBS, the adhesive SH‐SY5Y cells were observed by an inverted microscope (Nikon, Tokyo, Japan) and examined by MTT as described previously . The untreated SH‐SY5Y cells were used as the control group, and their absorbance at 490 nm (OD value) was described as 100%.…”

“…After SH-SY5Y cells were pretreated with Meri-olin1 (0.5, 1, 2 lM) for 48 h, 1 9 10 6 cells/ml of cell suspension was seeded into the precoated 96-well plate and incubated at 37°C for 2 h. When the unbound SH-SY5Y cells were washed twice with PBS, the adhesive SH-SY5Y cells were observed by an inverted microscope (Nikon, Tokyo, Japan) and examined by MTT as described previously. [25] The untreated SH-SY5Y cells were used as the control group, and their absorbance at 490 nm (OD value) was described as 100%. After treating SH-SY5Y cells with Meriolin1, the adhesion rate of SH-SY5Y cells was calculated according to the formula below: the adhesion rate of SH À SY5Y cells % of control ð Þ ¼ Meriolin1 treating groups'OD value/PBS treating group's OD value  100%.…”

Low concentrations of the recombinant toxin protein rLj-RGD3 suppress TNF-α-induced human renal carcinoma cell invasion

“…Our data indicated that rLj-RGD3 might initiate apoptosis in 786-0 cells by inducing actin filament aggregation. Our previous studies have shown that rLj-RGD3 can reduce the invasiveness of tumor cells (HeLa and MCF-7/Adr cells) and endothelial cells (ECV304 cells) [13,14]. However, the mechanisms by which rLj-RGD3 reduces cell invasion in vitro are largely uninvestigated.…”

“…Furthermore, rLj-RGD3 decreased the adhesion of tumor cells (HeLa and MCF-7/Adr cells) and endothelial cells (ECV304 cells) in vitro and inhibited avb3, avb5, and b1 integrin-mediated adhesion [13,14]. The mechanisms of rLj-RGD3 action included the inhibition of vitronectin or fibronectin-dependent tumor cell adhesion, destruction of the cytoskeleton, and blockade of tumor cell invasion through a reconstituted basement membrane (Matrigel) [13,14]. These findings led to further investigations about the functional effect of rLj-RGD3 on human carcinoma cell invasion in vitro.…”

“…We cloned the Lj-RGD3 gene, generated the recombinant protein rLj-RGD3, and results showed that rLj-RGD3 inhibited HeLa and MCF-7/Adr cell invasion and chicken chorioallantoic membrane angiogenesis. Furthermore, rLj-RGD3 decreased the adhesion of tumor cells (HeLa and MCF-7/Adr cells) and endothelial cells (ECV304 cells) in vitro and inhibited avb3, avb5, and b1 integrin-mediated adhesion [13,14]. The mechanisms of rLj-RGD3 action included the inhibition of vitronectin or fibronectin-dependent tumor cell adhesion, destruction of the cytoskeleton, and blockade of tumor cell invasion through a reconstituted basement membrane (Matrigel) [13,14].…”