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[Cell cycle arrest and apoptosis induced by tubeimoside in HeLa cell].

Ping Yang, Tingxi Yu, Rundi Ma, Li-Jian Yu, Weiming Su, Hai-yan Shao, Mingneng Liao, Huang Lai-zhen

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Abstract

BACKGROUND & OBJECTIVE: Tubeimoside, which is composed of tubeimoside I (79%) and II (21%), was isolated from the tubers of Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), a traditional Chinese medicine, "Tu-Bei-Mu". This study was designed to investigate the anti-tumor mechanism of tubeimoside. METHODS: Growth inhibition was measured by MTT assay. Induction of cell cycle arrest and apoptosis was determined by flow cytometry, fluorescence and electron microscopy, and gel electrophoresis of fragmented DNA. RESULTS: Tubeimoside display strong growth inhibitory effect in a dose- and time-dependant manner against HeLa cells with estimated IC50 values of 20.0, 18.8, and 8.8 mumol/L after 24, 48, and 72 h of treatment with tubeimoside. The flow cytometry profiles revealed that treatment with tubeimoside (5 h; 15, 30, 35 mumol/L) led to a dose-dependant shift from 9.80% up to 21.90%, and 27.00% in percentage of cells with a G2/M-like DNA content. On the other hand, treatment with tubeimoside (12 h, 15, 30, 35 mumol/L) led to a time-dependant shift from 8.20% up to 21.40%, 31.15%, and 34.55%, respectively. Exposure of HeLa cells to 40 mumol/L of tubeimoside induced nuclear shrinkage, chromation condensation and margination against nuclear envelope, subdiploid peak, and DNA fragmentation, characteristic as seen in apoptotic cells. CONCLUSION: Induction of cell cycle arrest and apoptosis may play an important role in the anti-tumor effect of tubeimoside.

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BACKGROUND & OBJECTIVE: Tubeimoside, which is composed of tubeimoside I (79%) and II (21%), was isolated from the tubers of Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), a traditional Chinese medicine, "Tu-Bei-Mu". This study was designed to investigate the anti-tumor mechanism of tubeimoside. METHODS: Growth inhibition was measured by MTT assay. Induction of cell cycle arrest and apoptosis was determined by flow cytometry, fluorescence and electron microscopy, and gel electrophoresis of fragmented DNA. RESULTS: Tubeimoside display strong growth inhibitory effect in a dose- and time-dependant manner against HeLa cells with estimated IC50 values of 20.0, 18.8, and 8.8 mumol/L after 24, 48, and 72 h of treatment with tubeimoside. The flow cytometry profiles revealed that treatment with tubeimoside (5 h; 15, 30, 35 mumol/L) led to a dose-dependant shift from 9.80% up to 21.90%, and 27.00% in percentage of cells with a G2/M-like DNA content. On the other hand, treatment with tubeimoside (12 h, 15, 30, 35 mumol/L) led to a time-dependant shift from 8.20% up to 21.40%, 31.15%, and 34.55%, respectively. Exposure of HeLa cells to 40 mumol/L of tubeimoside induced nuclear shrinkage, chromation condensation and margination against nuclear envelope, subdiploid peak, and DNA fragmentation, characteristic as seen in apoptotic cells. CONCLUSION: Induction of cell cycle arrest and apoptosis may play an important role in the anti-tumor effect of tubeimoside.

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Available abstract

BACKGROUND & OBJECTIVE: Tubeimoside, which is composed of tubeimoside I (79%) and II (21%), was isolated from the tubers of Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae), a traditional Chinese medicine, "Tu-Bei-Mu". This study was designed to investigate the anti-tumor mechanism of tubeimoside. METHODS: Growth inhibition was measured by MTT assay. Induction of cell cycle arrest and apoptosis was determined by flow cytometry, fluorescence and electron microscopy, and gel electrophoresis of fragmented DNA. RESULTS: Tubeimoside display strong growth inhibitory effect in a dose- and time-dependant manner against HeLa cells with estimated IC50 values of 20.0, 18.8, and 8.8 mumol/L after 24, 48, and 72 h of treatment with tubeimoside. The flow cytometry profiles revealed that treatment with tubeimoside (5 h; 15, 30, 35 mumol/L) led to a dose-dependant shift from 9.80% up to 21.90%, and 27.00% in percentage of cells with a G2/M-like DNA content. On the other hand, treatment with tubeimoside (12 h, 15, 30, 35 mumol/L) led to a time-dependant shift from 8.20% up to 21.40%, 31.15%, and 34.55%, respectively. Exposure of HeLa cells to 40 mumol/L of tubeimoside induced nuclear shrinkage, chromation condensation and margination against nuclear envelope, subdiploid peak, and DNA fragmentation, characteristic as seen in apoptotic cells. CONCLUSION: Induction of cell cycle arrest and apoptosis may play an important role in the anti-tumor effect of tubeimoside.

Key concepts: Apoptosis, HeLa, Flow cytometry, DNA fragmentation, Cell cycle, Cell cycle checkpoint, Fragmentation (computing), Cell growth

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