Component Analysis and Anti-Colorectal Cancer Mechanism via AKT/mTOR Signalling Pathway of Sanghuangporus vaninii Extracts
Abstract
:1. Introduction
2. Results
2.1. Total Polysaccharide and Flavonoids Contents
2.2. UPLC/Q-TOF-MS Analysis
2.3. The Result of CCK8 Cell Proliferation
2.4. Induction of Apoptosis by the Ethanol Extracts of S. vaninii
2.5. Effect of SVE60 on Cell Cycle Progression
2.6. The Result of qRT-PCR
2.7. The Result of SVE60 on the Protein Expression via mTOR Signalling Pathway
3. Materials and Methods
3.1. Materials
3.2. Preparation of the Phellinus Linteus Extracts
3.3. Total Polysaccharide Content
3.4. Total Flavonoids Content
3.5. UPLC-Q-TOF-MS Analytical Conditions
3.6. Cell Culture
3.7. CCK-8 Assay
3.8. Flow Cytometry Cell Apoptosis Analysis
3.9. Cell Cycle Analysis
3.10. Quantitative RT-PCR
3.11. Western Blot
3.12. Statistical Analysis
4. Conclusions and Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Peak No. | tR (min) | Identification | Formula | Mass (m/z) | Cacl. Mass (m/z) | mDa | Fragments |
---|---|---|---|---|---|---|---|
1 | 0.50 | 2-Carboxylbenzaldehyde | C8H6O3 | 151.0358 | 151.0395 | −3.7 | 151,128,110 |
2 | 0.55 | 8-hydroxyl-5-O-β-D-Glucopyranosylpsoralen | C17H16O10 | 381.0803 | 381.0822 | −1.9 | 381,365,353,258,104 |
3 | 0.55 | 7-(α-D-Glucopyranosyloxy)-2,3,4,5,6-pentahydroxyheptanoic acid | C13H24O13 | 387.1141 | 387.1139 | 0.2 | 387,341,245,181,129 |
4 | 0.62 | 2,3,4,5-Tetra-O-acetylhexonic acid | C14H20O11 | 365.1058 | 365.1084 | −2.6 | 365,229,205,175,124 |
5 | 0.75 | Adenosine | C10H13O4N5 | 268.1047 | 268.1046 | 0.1 | 268,245,229,136,124 |
6 | 0.75 | Citric acid | C6H8O7 | 191.0190 | 191.0192 | −0.2 | 191,173,128,111 |
7 | 1.34 | Protocatechuic acid | C7H6O4 | 153.0183 | 153.0188 | −0.5 | 153,109 |
8 | 2.01 | Protocatechuic aldehyde | C7H6O3 | 137.0235 | 137.0239 | −0.4 | 137,136 |
9 | 2.57 | Caffeic acid | C9H8O4 | 179.0342 | 179.0344 | −0.2 | 179,151,135,113 |
10 | 2.99 | Ethyl 6-hydroxy-1-cyclohexene-1-carboxylate | C9H14O3 | 171.0999 | 171.1021 | −2.2 | 229,171,158,138 |
11 | 3.78 | 2-(2-{2-[2-(2-Methoxyphenoxy) ethoxy]ethoxy} Ethoxy)ethanol | C15H24O6 | 299.1491 | 299.1495 | −0.4 | 299,249,207,147,113 |
12 | 4.18 | Osmundacetone | C10H10O3 | 177.0551 | 177.0552 | −0.1 | 177,161,133 |
13 | 4.89 | Hispidin | C13H10O5 | 245.0449 | 245.045 | −0.1 | 245,159,113 |
14 | 5.33 | Sternbin | C16H14O6 | 301.0706 | 301.0712 | −0.6 | 301,257,249,179,113 |
15 | 5.55 | 2,6-bis[3-(3-tert-butyl-2-hydroxy-5-methylphenyl)-3-tricyclo[5.2.1.02,6]decanyl]-4-methylphenol | C49H64O3 | 701.4937 | 701.4934 | 0.3 | 701,680,340,229,138 |
16 | 5.80 | Phelligridimer A or isomer | C52H32O20 | 977.1552 | 977.1565 | −1.3 | 977,301,245,229,142 |
17 | 5.86 | 4,4′-[2,7-Naphthalenediylbis(oxy)] diphthalic acid | C26H16O10 | 487.0648 | 487.0665 | −1.7 | 487,463,259,181,113 |
18 | 6.22 | Davallialactone | C25H20O9 | 463.1021 | 463.1029 | −0.8 | 463,379,259,159,113 |
19 | 6.29 | Phelligridimer A or isomer | C52H32O20 | 977.1552 | 977.1565 | −1.3 | 977,301,245,229,142 |
20 | 6.88 | 4-dimethyl methoxyphenylmethylene malonate | C13H14O5 | 249.0758 | 249.0763 | −0.5 | 249,219,159,113 |
21 | 7.00 | Unknown | C49H78O18 | 955.5246 | 955.5266 | −2.0 | 956,423,301,229,149 |
22 | 7.12 | Hosenkoside C | C48H82O20 | 977.5359 | 977.5321 | 3.8 | 978,932,113 |
23 | 7.89 | Hypholomine B | C26H18O10 | 491.0981 | 491.0978 | 0.3 | 491.301,183,142 |
8.01 | Hypholomine B | C26H18O10 | 489.0830 | 489.0822 | 0.8 | 489,445,199,147,113 | |
24 | 8.13 | Acetyl-SSa | C44H70O14 | 823.4819 | 823.4844 | −2.5 | 823,423,203,147,138 |
25 | 8.24 | 12-O-Acetylpergularin3-O-[β-D-oleandropyranosyl-(1→4)-β-D-canaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside] | C50H80O18 | 969.5384 | 969.5423 | −3.9 | 970,423,301,229,149 |
26 | 8.30 | Unknown | C36H78O21 | 845.4923 | 845.4957 | −3.4 | 845,445,249,130,113 |
27 | 8.43 | Muricatin II | C49H84O20 | 991.5508 | 991.5478 | 3.0 | 992,946,113 |
28 | 10.43 | Inoseavin A | C25H18O9 | 461.0863 | 461.0873 | −1.0 | 461,377,159,135,113 |
29 | 14.01 | Acetyl-SSa | C44H70O14 | 823.4802 | 823.4844 | −4.2 | 823,801,301,229,142 |
30 | 14.15 | Unknown | C36H78O21 | 845.4937 | 845.4957 | −2.0 | 845,799,113 |
31 | 14.99 | (3β,16β,24S)-cycloartane-3,16,24,25,30-pentol 3,25-di-β-D-glucopyranoside | C42H72O15 | 815.4830 | 815.4793 | 3.7 | 815,363,249,175,113 |
32 | 16.37 | Unknown | C61H66O2 | 829.4944 | 829.4985 | −4.1 | 829,786,385,147,113 |
33 | 17.26 | 5′,8′-dihydroxy-5,8-dimethoxy-6,6′-dimethyl-7,3′-binaphthyl- 1,4,1′,4′-tetraone | C24H18O8 | 433.0908 | 433.0923 | −1.5 | 433,385,179,147,113 |
34 | 17.36 | Unknown | C56H90O23 | 1131.5933 | 1131.5951 | −1.8 | 1,131,407,229,138 |
35 | 17.48 | Unknown | C48H98O30 | 1153.6039 | 1153.6065 | −2.6 | 1,154,599,489,113 |
36 | 17.79 | Chakasaponin VI | C59H92O26 | 1217.5917 | 1217.5955 | −3.8 | 1,218,301,229,138 |
37 | 18.14 | Unknown | C41H80O24 | 955.4695 | 955.4961 | 0.4 | 955,500,334,207,113 |
38 | 20.11 | 12-O-Acetyllineolon3-O-[β-D-oleandropyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside] | C50H80O18 | 969.5378 | 969.5423 | −4.1 | 970,767,425,229,149 |
39 | 20.20 | Muricatin IV | C49H84O20 | 991.5518 | 991.5478 | 4.0 | 992,946,113 |
40 | 20.42 | Cladoloside A4 | C53H82O21 | 1055.5393 | 1055.5427 | −3.4 | 1,056,875,301,229 |
41 | 20.50 | Unknown | C44H88O26 | 1031.5485 | 1031.5486 | −0.1 | 1,032,992,207,113 |
42 | 21.20 | 3β-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl]-olean-12-en-28-O-[(3-O-acetyl)-α-L-rhamnopyranosyl] ester | C50H80O18 | 969.5368 | 969.5364 | 0.4 | 970,407,301,229,138 |
43 | 21.29 | Merremoside c | C49H84O20 | 991.5511 | 991.5478 | 3.3 | 992,946,179,113 |
Apoptosis Rate | SVE60 | SVE95 | ||||||
---|---|---|---|---|---|---|---|---|
Control | 3 μg/mL | 30 μg/mL | 300 μg/mL | Control | 3 μg/mL | 30 μg/mL | 300 μg/mL | |
Total | 0.8% | 2.8% | 3.0% | 8.3% | 0.9% | 1.9% | 2.6% | 8.2% |
Early | 0.7% | 2.3% | 2.7% | 7.7% | 0.8% | 2.3% | 2.7% | 7.7% |
Late | 0.1% | 0.5% | 0.3% | 0.6% | 0.1% | 0.3% | 0.4% | 0.6% |
Name | Forward Primer (5′→3′) | Reverse Primer (5′→3′) |
---|---|---|
AKT | ATGAACGACGTAGCCATTGTG | TTGTAGCCAATAAAGGTGCCAT |
mTOR | ACCGGCACACATTTGAAGAAG | CTCGTTGAGGATCAGCAAGG |
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Guo, S.; Duan, W.; Wang, Y.; Chen, L.; Yang, C.; Gu, X.; Xue, Q.; Li, R.; Zhang, Z. Component Analysis and Anti-Colorectal Cancer Mechanism via AKT/mTOR Signalling Pathway of Sanghuangporus vaninii Extracts. Molecules 2022, 27, 1153. https://doi.org/10.3390/molecules27041153
Guo S, Duan W, Wang Y, Chen L, Yang C, Gu X, Xue Q, Li R, Zhang Z. Component Analysis and Anti-Colorectal Cancer Mechanism via AKT/mTOR Signalling Pathway of Sanghuangporus vaninii Extracts. Molecules. 2022; 27(4):1153. https://doi.org/10.3390/molecules27041153
Chicago/Turabian StyleGuo, Shanshan, Wenwen Duan, Yaxin Wang, Liangmian Chen, Chenchen Yang, Xuezhu Gu, Qinghai Xue, Raorao Li, and Zhijie Zhang. 2022. "Component Analysis and Anti-Colorectal Cancer Mechanism via AKT/mTOR Signalling Pathway of Sanghuangporus vaninii Extracts" Molecules 27, no. 4: 1153. https://doi.org/10.3390/molecules27041153