Related Experiment Video
Updated: Jul 8, 2026
Colony Formation Assay Detecting the Proliferative Capacity of LncRNA-knockdown Osteosarcoma Cells
Published on: January 16, 2026
[Construction and screening of cDNA library of osteosarcoma cells]
Shao-lin Deng1, Yi Quan, Xian-ming Pan
1General Hospital of Chengdu Command, Chengdu 610083, China. deng shaolin@medmail.com.cn
Objective:
To construct a cDNA library of osteosarcoma SAOS-2 cells and screen the proteins interacting with the bone morphogenetic protein-2 (BMP-2) to explore the role of BMP-2 in the development of prostate cancer.
Methods:
The total RNA was extracted from SAOS-2 cells, from which poly(A)(+)RNA was purified to generate the cDNA using RT-PCR with primer SMARTIII and primer CDSIIIoligo(dT). The library was constructed and screened by cotransformation of the double-stranded cDNA (dscDNA) and linearized pGADT7-Rec with the bait plasmid to the yeast AH109. The proteins interacting with BMP-2 were screened by AH109 mating with the bait strain Y187, and the interaction was confirmed using far-Western blot analysis.
Results:
The cDNA library of human osteosarcoma SAOS-2 cells was constructed, which was characterized by high diversity and sufficient capacity. The dscDNA size range was between 250-5000 bp, with a cotransformation efficiency of 4.3 x 10(5) and recombination efficiency of 1.9 x 10(6), and 4.3 x 10(5) clones were screened. The mating efficiency was 32% and 1.0 x 10(6) clones were screened by the bait of BMP-2, and 4 positive clones was obtained and sequenced.
Conclusion:
The diversity and capacity of the cDNA library meet the needs for screening genes related to prostate cancer.
Insights
A novel cDNA library from osteosarcoma cells was created to identify proteins interacting with bone morphogenetic protein-2 (BMP-2). This research aims to uncover BMP-2’s role in prostate cancer development.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Context:
- Osteosarcoma SAOS-2 cells provide a source for constructing a diverse cDNA library.
- Bone morphogenetic protein-2 (BMP-2) is implicated in various cellular processes, including cancer development.
- Understanding BMP-2 interactions is crucial for elucidating its role in prostate cancer.
Purpose:
- To construct a high-diversity cDNA library from human osteosarcoma SAOS-2 cells.
- To screen for proteins that interact with bone morphogenetic protein-2 (BMP-2).
- To investigate the potential role of BMP-2 in prostate cancer pathogenesis.
Summary:
- A cDNA library was successfully constructed from SAOS-2 cells with high diversity and capacity.
- Yeast-based screening identified four positive clones interacting with BMP-2.
- Far-Western blot analysis confirmed the interactions, validating the library's utility.
Impact:
- The constructed cDNA library meets the requirements for screening prostate cancer-related genes.
- This resource facilitates further investigation into BMP-2 signaling pathways in cancer.
- Findings may contribute to developing novel therapeutic strategies for prostate cancer.
