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Knockdown of Ski decreases osteosarcoma cell proliferation and migration by suppressing the PI3K/Akt signaling
Xin Zhao1, Yuying Fang2, Xingwen Wang3
1Department of Orthopedic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Abstract:
Ski, an evolutionary conserved protein, is involved in the development of a number of tumors, such as Barrett's esophagus, leukemia, colorectal cancer, gastric cancer, pancreatic cancer, hemangiomas and melanoma. However, studies on the functions of Ski in osteosarcoma (OS) are limited. In this study, firstly the differential expression of Ski in OS tissues and osteochondroma tissues was detected, and the expression of Ski in both human OS cell lines (MG63 and U2OS) and normal osteoblasts (hFoB1.19) was then detected. The results demonstrated that Ski expression was significantly upregulated in both human OS tissues and cell lines. The results led us to hypothesize that Ski may play an essential role in the pathological process of OS. Thus, Ski specific small interfere RNA (Ski‑siRNA) was used. The results revealed that OS cell proliferation was markedly inhibited following the knockdown of Ski, which was identified by CCK8 assay, EdU staining and cell cycle analysis. In addition, OS cell migration was significantly suppressed following Ski knockdown, which was identified by wound healing assay. Moreover, the protein levels of p‑PI3K and p‑Akt in OS cells declined prominently following Ski knockdown. On the whole, the findings of this study revealed that Ski expression was significantly upregulated in OS tissue and OS cells. The knockdown of Ski decreased OS cell proliferation and migration, which was mediated by blocking the PI3K/Akt signaling pathway. Thus, Ski may act as a tumor promoter gene in tumorigenesis, and Ski may prove to be a potential therapeutic target for the treatment of OS.
Insights
Ski protein is upregulated in osteosarcoma (OS) and drives cancer cell proliferation and migration. Inhibiting Ski may offer a new therapeutic strategy for treating OS by blocking the PI3K/Akt pathway.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Ski is an evolutionarily conserved protein implicated in various cancers.
- Limited research exists on Ski's role in osteosarcoma (OS) development.
Purpose of the Study:
- To investigate the differential expression of Ski in OS.
- To elucidate the functional role of Ski in OS cell proliferation and migration.
- To explore the underlying molecular mechanisms, including the PI3K/Akt signaling pathway.
Main Methods:
- Differential expression analysis of Ski in OS tissues and cell lines.
- Ski knockdown using small interfering RNA (siRNA).
- Cell proliferation assays (CCK8, EdU staining, cell cycle analysis).
- Cell migration assay (wound healing).
- Western blot analysis for PI3K/Akt pathway proteins.
Main Results:
- Ski expression was significantly upregulated in OS tissues and cell lines compared to normal tissues and cells.
- Ski knockdown markedly inhibited OS cell proliferation and migration.
- Ski knockdown led to a prominent decrease in the phosphorylation of PI3K and Akt proteins.
Conclusions:
- Ski acts as a tumor promoter in osteosarcoma by enhancing cell proliferation and migration.
- Ski's oncogenic function in OS is mediated through the PI3K/Akt signaling pathway.
- Ski represents a potential therapeutic target for osteosarcoma treatment.
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