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Integr Cancer Ther
2020 Jan 01;19:1534735420911430. doi: 10.1177/1534735420911430.
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Effects of Holothurian Glycosaminoglycan on the Sensitivity of Lung Cancer to Chemotherapy.
Lin C
,
Zhu X
,
Jin Q
,
Sui A
,
Li J
,
Shen L
.
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Sea cucumber is a kind of food. Holothurian glycosaminoglycan (hGAG) is extracted from the body wall of the sea cucumber. Administration of hGAG and cisplatin (DDP) together to treat lung cancer was investigated. Lung adenocarcinoma A549 cells were cultured and divided into 4 groups: control group, hGAG 100 µg/mL group, DDP 3 µg/mL group, and hGAG 100 µg/mL + DDP 3 µg/mL group. Cell inhibition and apoptosis was evaluated by CCK8 and Hoechst33258 staining. Cell cycle was tested by Annexin V-FITC/PI (propidium iodide) double-staining and flow cytometry. The expression of mRNA and protein of Bcl-2, Bax, caspase-3, and survivin were detected by reverse transcriptase-polymerase chain reaction and Western blot, respectively. The results showed that hGAG combined with DDP enhanced the inhibitory effect of DDP on A549 lung cells through apoptosis pathway. The mechanism of apoptosis may be related to the reduction of Bcl-2 and survivin, as well as the ascension of Bax and caspase-3. hGAG could promote A549 cell cycle arrest in G1 and G2 phase and improve the DDP chemotherapy effects on A549 cells.
Figure 1. A549 cells were treated with various concentrations of samples for 48 hours. The cells stained by Hochest33258 and images were captured using a fluorescence microscope. White arrow showed the apoptotic, shrunken, hyperchromatic, and pyknotic apoptotic cells.
Figure 2. A549 cells were treated with various concentrations of samples for 48 hours. Cell apoptosis tested by Annexin V-FITC/PI flow cytometry; proportion of apoptotic cells was measured.
Figure 3. A549 cells were treated with various concentrations of samples for 36 hours. The mRNA expression of Bcl-2, Bax, survivin, and caspase-3 in different groups. *P < .05 compared with the control group; **P < .05 hGAG + DDP group compared with DDP group.
Figure 4. A549 cells were treated with various concentrations of samples for 48 hours. The protein expression of Bcl-2, Bax, survivin, and caspase-3 in A549 cells detected by Western blotting in different groups. *P < .05 compared with the control group; **P < .05 hGAG + DDP group compared with DDP group.
Figure 5. A549 cells were treated with various concentrations of samples for 48 hours. Cell cycle changes and the distribution of each cell cycle phase in A549 cells analyzed by flow cytometry assays.
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