Colony Formation Assay Detecting the Proliferative Capacity of LncRNA-knockdown Osteosarcoma Cells

Aberrant long non-coding RNAs (lncRNAs) expression profoundly influences cellular proliferation of osteosarcoma cells, suggesting that they may serve as potential therapeutic targets. However, functional studies of lncRNAs remain largely theoretical and limited to a few validations. Colony formation assays, which specifically reflect the ultimate proliferative fate of single cells, are regarded as the gold standard for evaluating long-term proliferative potential. Here, this protocol established a reliable experimental protocol using colony formation assays in lncRNA-knockdown cells to evaluate the effect of lncRNA on osteosarcoma cell proliferative capacity. This experimental protocol provides a stable, cost-effective, and efficient assay to assess the regulatory effects of lncRNA on osteosarcoma cell proliferation. This approach is not only suitable for research on osteosarcoma cells but also serves as a feasible tool for lncRNA-regulated cell proliferative capacity in other tumors. This study included a detailed detection protocol of lncRNA-knockdown and colony formation assays, with an example of the small nucleolar RNA host gene 6 (SNHG6) knockdown in osteosarcoma cells (143B). Experimental results confirmed efficient suppression of SNHG6 expression, accompanied by a marked reduction in both the number and size of colonies. These findings suggest that SNHG6 is essential for sustaining the long-term proliferative potential of osteosarcoma cells (143B).