Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells
Abstract
:1. Introduction
2. Results
2.1. Growth Inhibitory Effects of LcS and Geniposide in HOK and HSC-3 Cells
2.2. Sub-G1 Content of HSC-3 Cells
2.3. mRNA and Protein Expressions of Caspase-3, Caspase-8 and Caspase-9 in HSC-3 Cells
2.4. mRNA and Protein Expressions of Bax, Bcl-2 and Bcl-xL in HSC-3 Cells
2.5. mRNA and Protein Expressions of p53 and p21 in HSC-3 Cells
2.6. mRNA and Protein Expressions of HIAP-1 and HIAP-2 in HSC-3 Cells
2.7. mRNA and Protein Expressions of NF-κB and IκB-α in HSC-3 Cells
2.8. mRNA and Protein Expressions of Fas and FasL in HSC-3 Cells
2.9. mRNA and Protein Expressions of TIMP-1, TIMP-2, MMP-2 and MMP-9 in HSC-3 Cells
2.10. mRNA and Protein Expressions of COX-2 and iNOS in HSC-3 Cells
2.11. LcS TransformsGeniposide to Genipin
2.12. Enzyme Activity of LcS Produce β-Glucosidase
3. Discussion
4. Materials and Methods
4.1. Preparation of the Experimental Sample
4.2. Cell Lines
4.3. Preparation for the Combination of Geniposide and LcS
4.4. MTT Assay
4.5. Flow Cytometry Assay
4.6. Real-Time Quantitative PCR Assay
4.7. Western Blot Assay
4.8. Liquid Chromatography Experiment
4.9. β-glucosidase Determination
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are not available from the authors. |
Treatment | OD570 Value | Inhibitory Rate (%) |
---|---|---|
Control | 0.479 ± 0.007 | / |
LcS | 0.454 ± 0.005 * | 5.2 ± 0.3 |
Geniposide-L | 0.365 ± 0.010 * | 23.8 ± 2.6 |
LcS-geniposide-L | 0.286 ± 0.009 * | 40.3 ± 2.3 |
Geniposide-H | 0.212 ± 0.008 ** | 55.7 ± 2.1 |
LcS-geniposide-H | 0.162 ± 0.005 ** | 73.7 ± 1.8 |
Gene Name | Sequence |
---|---|
Caspase-3 | Forward: 5′-CAA ACT TTT TCA GAG GGG ATC G-3′ |
Reverse: 5′-GCA TAC TGT TTC AGC ATG GCA-3′ | |
Caspase-8 | Forward: 5′-CCC CAC CCT CAC TTT GCT-3′ |
Reverse: 5′-GGA GGA CCA GGC TCA CTT A-3′ | |
Caspase-9 | Forward: 5′-GGC CCT TCC TCG CTT CAT CTC-3′ |
Reverse: 5′-GGT CCT TGG GCC TTC CTG GTA T-3′ | |
Bax | Forward: 5′-AAG CTG AGC GAG TGT CTC CGG CG-3′ |
Reverse: 5′-CAG ATG CCG GTT CAG GTA CTC AGT C-3′ | |
Bcl-2 | Forward: 5′-CTC GTC GCT ACC GTC GTG ACT TGG-3′ |
Reverse: 5′-CAG ATG CCG GTT CAG GTA CTC AGT C-3′ | |
Bcl-xL | Forward: 5′-CCC AGA AAG GAT ACA GCT GG-3′ |
Reverse: 5′-GCG ATC CGA CTC ACC AAT AC-3′ | |
Fas | Forward: 5′-GAA ATG AAA TCC AAA GCT-3′ |
Reverse: 5′-TAA TTT AGA GGC AAA GTG GC-3′ | |
FasL | Forward: 5′-GGA TTG GGC CTG GGG ATG TTT CA-3′ |
Reverse: 5′-TTG TGG CTC AGG GGC AGG TTG TTG-3′ | |
p53 | Forward: 5′-GCT CTG ACT GTA CCA CCA TCC-3′ |
Reverse: 5′-CTC TCG GAA CAT CTC GAA GCG-3′ | |
p21 | Forward: 5′-CTC AGA GGA GGC GCC ATG-3′ |
Reverse: 5′-GGG CGG ATT AGG GCT TCC-3′ | |
HIAP-1 | Forward: 5′-GCC TGA TGC TGG ATA ACT GG-3′ |
Reverse: 5′-GGC GAC AGA AAA GTC AAT GG-3′ | |
HIAP-2 | Forward: 5′-GCC TGA TGC TGG ATA ACT GG-3′ |
Reverse: 5′-GCT CTT GCC AAT TCT GAT GG-3′ | |
NF-κB | Forward: 5′-CAC TTA TGG ACA ACT ATG AGG TCT CTG-3′ |
Reverse: 5′-CTG TCT TGT GGA CAA CGC AGT GGA ATT-3′ | |
IκB-α | Forward: 5′-GCT GAA GAA GGA GCG GCT ACT-3′ |
Reverse: 5′-TCG TAC TCC TCG TCT TTC ATG GA-3′ | |
COX-2 | Forward: 5′-TTA AAA TGA GAT TGT CCG AA-3′ |
Reverse: 5′-AGA TCA CCT CTG CCT GAG TA-3′ | |
iNOS | Forward: 5′-AGA GAG ATC GGG TTC ACA-3′ |
Reverse: 5′-CAC AGA ACT GAG GGT ACA-3′ | |
MMP-2 | Forward: 5′-CTT CTT CAA GGA CCG GTT CA-3′ |
Reverse: 5′-GCT GGC TGA GTA CCA GTA-3′ | |
MMP-9 | Forward: 5′-TGG GCT ACG TGA CCT ATG AC-3′ |
Reverse: 5′-GCC CAG CCC ACC TCC ACT CC-3′ | |
TIMP-1 | Forward: 5′-GTC AGT GAG AAG CAA GTC GA-3′ |
Reverse: 5′-ATG TTC TTC TCT GTG ACC CA-3′ | |
TIMP-2 | Forward: 5′-TGG GGA CAC CAG AAG TCA AC-3′ |
Reverse: 5′-TTT TCA GAG CCT TGG AGG AG-3′ | |
GAPDH | Forward: 5′-CGG AGT CAA CGG ATT TGG TC-3′ |
Reverse: 5′-AGC CTT CTC CAT GGT CGT GA-3′ |
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Qian, Y.; Song, J.-L.; Sun, P.; Yi, R.; Liu, H.; Feng, X.; Park, K.-Y.; Zhao, X. Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells. Molecules 2018, 23, 1069. https://doi.org/10.3390/molecules23051069
Qian Y, Song J-L, Sun P, Yi R, Liu H, Feng X, Park K-Y, Zhao X. Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells. Molecules. 2018; 23(5):1069. https://doi.org/10.3390/molecules23051069
Chicago/Turabian StyleQian, Yu, Jia-Le Song, Peng Sun, Ruokun Yi, Honglin Liu, Xia Feng, Kun-Young Park, and Xin Zhao. 2018. "Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells" Molecules 23, no. 5: 1069. https://doi.org/10.3390/molecules23051069