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Antiproliferation and cell apoptosis inducing bioactivities of constituents from Dysosma versipellis in PC3 and Bcap-37 cell lines.

Xu X, Gao X, Jin L, Bhadury PS, Yuan K, Hu D, Song B, Yang S - Cell Div (2011)

Bottom Line: The inhibitory activities of these compounds were investigated on tumor cells PC3, Bcap-37 and BGC-823 in vitro by MTT method, and the results showed that podophyllotoxone (PTO) and 4'-demethyldeoxypodophyllotoxin (DDPT) had potent inhibitory activities against the growth of human carcinoma cell lines.This study suggests that most of the compounds from the roots of D. versipellis could inhibit the growth of human carcinoma cells.In addition, PTO and DDPT could induce apoptosis of tumor cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China. songbaoan22@yahoo.com.

ABSTRACT

Background: Recently, interest in phytochemicals from traditional Chinese medicinal herbs with the capability to inhibit cancer cells growth and proliferation has been growing rapidly due to their nontoxic nature. Dysosma versipellis as Bereridaceae plants is an endemic species in China, which has been proved to be an important Chinese herbal medicine because of its biological activity. However, systematic and comprehensive studies on the phytochemicals from Dysosma versipellis and their bioactivity are limited.

Results: Fifteen compounds were isolated and characterized from the roots of Dysosma versipellis, among which six compounds were isolated from this plant for the first time. The inhibitory activities of these compounds were investigated on tumor cells PC3, Bcap-37 and BGC-823 in vitro by MTT method, and the results showed that podophyllotoxone (PTO) and 4'-demethyldeoxypodophyllotoxin (DDPT) had potent inhibitory activities against the growth of human carcinoma cell lines. Subsequent fluorescence staining and flow cytometry analysis indicated that these two compounds could induce apoptosis in PC3 and Bcap-37 cells, and the apoptosis ratios reached the peak (12.0% and 14.1%) after 72 h of treatment at 20 μM, respectively.

Conclusions: This study suggests that most of the compounds from the roots of D. versipellis could inhibit the growth of human carcinoma cells. In addition, PTO and DDPT could induce apoptosis of tumor cells.

No MeSH data available.


Related in: MedlinePlus

Nuclei morphological changes during PTO and DDPT-induced apoptosis in tumor cells detected by AO/EB staining. After treated with PTO and DDPT at 20 μM, PC3 and Bcap-37 cells were stained with AO/EB (100 μg/mL) and observed under fluorescence microscopy. For PC3 cells group, A: negative control (without treatment); D: positive control, treated with HCPT (20 μM) for 24 h; B and C: treated with PTO (20 μM) for 24, 48 h; E and F, treated with DDPT (20 μM) for 24, 48 h, respectively. For Bcap-37 cells group, A': negative control; D': positive control, treated by HCPT (20 μM) for 24 h; B' and C': treated with PTO (20 μM) for 24, 48 h; E'and F': treated with DDPT (20 μM) for 24, 48 h, respectively.
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Figure 4: Nuclei morphological changes during PTO and DDPT-induced apoptosis in tumor cells detected by AO/EB staining. After treated with PTO and DDPT at 20 μM, PC3 and Bcap-37 cells were stained with AO/EB (100 μg/mL) and observed under fluorescence microscopy. For PC3 cells group, A: negative control (without treatment); D: positive control, treated with HCPT (20 μM) for 24 h; B and C: treated with PTO (20 μM) for 24, 48 h; E and F, treated with DDPT (20 μM) for 24, 48 h, respectively. For Bcap-37 cells group, A': negative control; D': positive control, treated by HCPT (20 μM) for 24 h; B' and C': treated with PTO (20 μM) for 24, 48 h; E'and F': treated with DDPT (20 μM) for 24, 48 h, respectively.

Mentions: Since acridine orange (AO) was a vital dye that could stain nuclear DNA across an intact cell membrane and ethidium bromide (EB) could only stain cells that had lost membrane integrity. Thus, live cells will be uniformly stained green, early apoptotic cells will be densely stained as green yellow or displayed green yellow fragments, while late apoptotic cells will be densely stained as orange or displayed orange fragments, and necrotic cells will be stained with orange with no condensed chromatin could be found by the AO/EB doubly staining. After tumor cells were treated with PTO and DDPT (20 μM) for 24, 48 h, the morphological changes were analyzed. As shown in Figure 4, green live PC3 and Bcap-37 cells with normal morphology were seen in the negative control group (Figure 4A and 4A'). In contrast, early apoptotic cells with yellow green dots in PC3 cell nuclei and late apoptotic cells with orange dots in Bcap-37 cell nuclei could be seen in the positive control group (Figure 4D and 4D'), meanwhile bright green early apoptotic cells with nuclear margination and chromatin condensation occurred in the experimental group with 24 h of treatment (Figure 4B and 4E) and orange colored apoptotic cells with apoptotic bodies and chromatin fragmentation could be seen when PTO and DDPT were applied for 48 h (Figure 4E and 4F). Similarly, morphological changes of Bcap-37 cell apoptosis were also observed under microscope (Figure 4B', 4C', 4E', and 4F'). The results suggested that PTO and DDPT were able to induce apoptosis in PC3 and Bcap-37 cells.


Antiproliferation and cell apoptosis inducing bioactivities of constituents from Dysosma versipellis in PC3 and Bcap-37 cell lines.

Xu X, Gao X, Jin L, Bhadury PS, Yuan K, Hu D, Song B, Yang S - Cell Div (2011)

Nuclei morphological changes during PTO and DDPT-induced apoptosis in tumor cells detected by AO/EB staining. After treated with PTO and DDPT at 20 μM, PC3 and Bcap-37 cells were stained with AO/EB (100 μg/mL) and observed under fluorescence microscopy. For PC3 cells group, A: negative control (without treatment); D: positive control, treated with HCPT (20 μM) for 24 h; B and C: treated with PTO (20 μM) for 24, 48 h; E and F, treated with DDPT (20 μM) for 24, 48 h, respectively. For Bcap-37 cells group, A': negative control; D': positive control, treated by HCPT (20 μM) for 24 h; B' and C': treated with PTO (20 μM) for 24, 48 h; E'and F': treated with DDPT (20 μM) for 24, 48 h, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3141614&req=5

Figure 4: Nuclei morphological changes during PTO and DDPT-induced apoptosis in tumor cells detected by AO/EB staining. After treated with PTO and DDPT at 20 μM, PC3 and Bcap-37 cells were stained with AO/EB (100 μg/mL) and observed under fluorescence microscopy. For PC3 cells group, A: negative control (without treatment); D: positive control, treated with HCPT (20 μM) for 24 h; B and C: treated with PTO (20 μM) for 24, 48 h; E and F, treated with DDPT (20 μM) for 24, 48 h, respectively. For Bcap-37 cells group, A': negative control; D': positive control, treated by HCPT (20 μM) for 24 h; B' and C': treated with PTO (20 μM) for 24, 48 h; E'and F': treated with DDPT (20 μM) for 24, 48 h, respectively.
Mentions: Since acridine orange (AO) was a vital dye that could stain nuclear DNA across an intact cell membrane and ethidium bromide (EB) could only stain cells that had lost membrane integrity. Thus, live cells will be uniformly stained green, early apoptotic cells will be densely stained as green yellow or displayed green yellow fragments, while late apoptotic cells will be densely stained as orange or displayed orange fragments, and necrotic cells will be stained with orange with no condensed chromatin could be found by the AO/EB doubly staining. After tumor cells were treated with PTO and DDPT (20 μM) for 24, 48 h, the morphological changes were analyzed. As shown in Figure 4, green live PC3 and Bcap-37 cells with normal morphology were seen in the negative control group (Figure 4A and 4A'). In contrast, early apoptotic cells with yellow green dots in PC3 cell nuclei and late apoptotic cells with orange dots in Bcap-37 cell nuclei could be seen in the positive control group (Figure 4D and 4D'), meanwhile bright green early apoptotic cells with nuclear margination and chromatin condensation occurred in the experimental group with 24 h of treatment (Figure 4B and 4E) and orange colored apoptotic cells with apoptotic bodies and chromatin fragmentation could be seen when PTO and DDPT were applied for 48 h (Figure 4E and 4F). Similarly, morphological changes of Bcap-37 cell apoptosis were also observed under microscope (Figure 4B', 4C', 4E', and 4F'). The results suggested that PTO and DDPT were able to induce apoptosis in PC3 and Bcap-37 cells.

Bottom Line: The inhibitory activities of these compounds were investigated on tumor cells PC3, Bcap-37 and BGC-823 in vitro by MTT method, and the results showed that podophyllotoxone (PTO) and 4'-demethyldeoxypodophyllotoxin (DDPT) had potent inhibitory activities against the growth of human carcinoma cell lines.This study suggests that most of the compounds from the roots of D. versipellis could inhibit the growth of human carcinoma cells.In addition, PTO and DDPT could induce apoptosis of tumor cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China. songbaoan22@yahoo.com.

ABSTRACT

Background: Recently, interest in phytochemicals from traditional Chinese medicinal herbs with the capability to inhibit cancer cells growth and proliferation has been growing rapidly due to their nontoxic nature. Dysosma versipellis as Bereridaceae plants is an endemic species in China, which has been proved to be an important Chinese herbal medicine because of its biological activity. However, systematic and comprehensive studies on the phytochemicals from Dysosma versipellis and their bioactivity are limited.

Results: Fifteen compounds were isolated and characterized from the roots of Dysosma versipellis, among which six compounds were isolated from this plant for the first time. The inhibitory activities of these compounds were investigated on tumor cells PC3, Bcap-37 and BGC-823 in vitro by MTT method, and the results showed that podophyllotoxone (PTO) and 4'-demethyldeoxypodophyllotoxin (DDPT) had potent inhibitory activities against the growth of human carcinoma cell lines. Subsequent fluorescence staining and flow cytometry analysis indicated that these two compounds could induce apoptosis in PC3 and Bcap-37 cells, and the apoptosis ratios reached the peak (12.0% and 14.1%) after 72 h of treatment at 20 μM, respectively.

Conclusions: This study suggests that most of the compounds from the roots of D. versipellis could inhibit the growth of human carcinoma cells. In addition, PTO and DDPT could induce apoptosis of tumor cells.

No MeSH data available.


Related in: MedlinePlus