Limits...
Involvement of Seladin-1 in goniothalamin-induced apoptosis in urinary bladder cancer cells.

Yen HK, Fauzi AR, Din LB, McKelvey-Martin VJ, Meng CK, Inayat-Hussain SH, Rajab NF - BMC Complement Altern Med (2014)

Bottom Line: This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells.The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. nfadilah@medic.ukm.my.

ABSTRACT

Background: Selective Alzheimer Disease Indicator-1 (or Seladin-1) is a multifunctional protein first discovered by downregulation of its expression in Alzheimer's disease. Interestingly, the expression of this protein is upregulated in several cancers, including primary bladder cancer. However, its role in cancer formation has yet to be discovered. Goniothalamin is a natural product that has been demonstrated to induce apoptosis in various cancer cell lines. In this study, we have elucidated the role of Seladin-1 in goniothalamin-induced cytotoxicity towards human urinary bladder cancer cell line RT4.

Methods: The cytotoxicity of goniothalamin in human urinary bladder cancer cell line RT4 was assessed using MTT assay and the mode of cell death was determined by Annexin V-FITC/PI labeling assay. Finally, the expression of Seladin-1 protein in goniothalamin-treated RT4 cells was determined by Western blot.

Results: MTT assay showed that the cytotoxicity of goniothalamin on RT4 cells was concentration and time dependent with IC50 values of 61 μM (24 hr), 38 μM (48 hr) and 31 μM for 72 hr, respectively. Cell death induced was confirmed through apoptosis; as assessed using the Annexin V-FITC/PI labeling assay. Furthermore, the involvement of Seladin-1 in goniothalamin-induced apoptosis was evidenced through the cleavage of 60 kDa protein to 40 kDa and 20 kDa. This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.

Conclusion: This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells. The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

Show MeSH

Related in: MedlinePlus

Expression of Seladin-1 on RT4 cells after treatment by IC50(61 μM) of goniothalamin at 2, 4, 8, 16, and 24 hrs. Vehicle control (or untreated cells) acted as a negative control and β-actin as a loading control during the experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4150971&req=5

Fig4: Expression of Seladin-1 on RT4 cells after treatment by IC50(61 μM) of goniothalamin at 2, 4, 8, 16, and 24 hrs. Vehicle control (or untreated cells) acted as a negative control and β-actin as a loading control during the experiment.

Mentions: The involvement of Seladin-1 protein on induction by goniothalamin was assessed using Western blotting (as shown in Figure 4). RT4 cells were treated by IC50 of goniothalamin at 2, 4, 8, 16, and 24 hrs to assess the expression of Seladin-1. The results showed that there was a cleavage of Seladin-1 from 60 kDa to 40 kDa, and 20 kDa as increase in treatment time, and the 40 kDa fragment was gradually decreased but not visualized at a later time point. On the contrary, the 20 kDa fragment was gradually increased from 8, 16, and 24 hours.Figure 4


Involvement of Seladin-1 in goniothalamin-induced apoptosis in urinary bladder cancer cells.

Yen HK, Fauzi AR, Din LB, McKelvey-Martin VJ, Meng CK, Inayat-Hussain SH, Rajab NF - BMC Complement Altern Med (2014)

Expression of Seladin-1 on RT4 cells after treatment by IC50(61 μM) of goniothalamin at 2, 4, 8, 16, and 24 hrs. Vehicle control (or untreated cells) acted as a negative control and β-actin as a loading control during the experiment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150971&req=5

Fig4: Expression of Seladin-1 on RT4 cells after treatment by IC50(61 μM) of goniothalamin at 2, 4, 8, 16, and 24 hrs. Vehicle control (or untreated cells) acted as a negative control and β-actin as a loading control during the experiment.
Mentions: The involvement of Seladin-1 protein on induction by goniothalamin was assessed using Western blotting (as shown in Figure 4). RT4 cells were treated by IC50 of goniothalamin at 2, 4, 8, 16, and 24 hrs to assess the expression of Seladin-1. The results showed that there was a cleavage of Seladin-1 from 60 kDa to 40 kDa, and 20 kDa as increase in treatment time, and the 40 kDa fragment was gradually decreased but not visualized at a later time point. On the contrary, the 20 kDa fragment was gradually increased from 8, 16, and 24 hours.Figure 4

Bottom Line: This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells.The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. nfadilah@medic.ukm.my.

ABSTRACT

Background: Selective Alzheimer Disease Indicator-1 (or Seladin-1) is a multifunctional protein first discovered by downregulation of its expression in Alzheimer's disease. Interestingly, the expression of this protein is upregulated in several cancers, including primary bladder cancer. However, its role in cancer formation has yet to be discovered. Goniothalamin is a natural product that has been demonstrated to induce apoptosis in various cancer cell lines. In this study, we have elucidated the role of Seladin-1 in goniothalamin-induced cytotoxicity towards human urinary bladder cancer cell line RT4.

Methods: The cytotoxicity of goniothalamin in human urinary bladder cancer cell line RT4 was assessed using MTT assay and the mode of cell death was determined by Annexin V-FITC/PI labeling assay. Finally, the expression of Seladin-1 protein in goniothalamin-treated RT4 cells was determined by Western blot.

Results: MTT assay showed that the cytotoxicity of goniothalamin on RT4 cells was concentration and time dependent with IC50 values of 61 μM (24 hr), 38 μM (48 hr) and 31 μM for 72 hr, respectively. Cell death induced was confirmed through apoptosis; as assessed using the Annexin V-FITC/PI labeling assay. Furthermore, the involvement of Seladin-1 in goniothalamin-induced apoptosis was evidenced through the cleavage of 60 kDa protein to 40 kDa and 20 kDa. This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.

Conclusion: This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells. The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

Show MeSH
Related in: MedlinePlus