Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on lung cancer cell proliferation. Cell viability of A549 ( A ), A549-TR ( B ), and H1299 ( C ) was determined by MTT assay after treatment with MH for 72 h. Data represent mean ± S.D. of 3 independent experiments. MH dose-dependently inhibited the colony-forming ability of NSCLC cells. ( D ) Images depict the colony-forming assay of drug-sensitive (A549 and H1299) and drug-resistant (A549-TR) NSCLC cells. Bar diagrams represent the number of colonies of NSCLC cells treated with different doses of MH ( E – G ). Statistical analysis was performed using Student t -test with Graphpad Prism version 5 software. **, p < 0.01; ***, p < 0.001.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: MTT Assay, Software

Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on cell cycle markers in lung cancer cells. ( A ) A schematic diagram illustrating MH’s effects on various phases of the cell cycle arresting the G 0 /G 1 and G 2 phases. Following 48 h MH treatment, protein lysates from NSCLC cells were analyzed by Western blot for cell cycle regulatory markers in A549 ( B ), A549-TR ( C ), and H1299 ( D ) cells. β-Actin was used as loading control. Numbers above the band depict relative expression (RE) compared to vehicle treatment.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: Western Blot, Control, Expressing

Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on apoptotic markers in A549 ( A ), A549-TR ( B ), and H1299 ( C ) cells. NSCLC cells were treated with MH for 48 h, and protein lysates were probed to assess the levels of apoptosis markers. β-Actin was used for equal loading. Numbers above each band depict relative expression (RE) compared to vehicle-treated cells. and share the same β-Actin blot for BCL-2 and Survivin in A549-TR cells.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: Expressing

Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on the MET/PI3K/Akt/mTOR pathway. MH induces apoptosis in NSCLC cells by blocking the MET/AKT/mTOR/survivin signaling pathway. The impact of MH treatment on oncogene expression in NSCLC cells A549 ( A ), A549-TR ( B ), and H1299 ( C ) after 48 h is shown through immunoblotting analysis. Equal loading is represented by the levels of β-Actin. Numbers above the band depict the relative expression (RE) in comparison with vehicle-treated cells. , and share the same β-Actin for a few protein markers.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: Blocking Assay, Expressing, Western Blot, Comparison

Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on RAS/cMYC and NF-kB. Cells were treated with MH for 48 h, and whole cell and nuclear lysates were prepared. The effect of MH on RAS/cMYC using whole cell lysates and NF-κB in nuclear extracts of NSCLC A549 ( A ), A549-TR ( B ), and H1299 ( C ) cells was determined by Western blot analysis. Equal loading was determined by the levels of β-Actin (whole cell lysates) and Lamin B1 (nuclear extracts). Numbers above each band depict the relative expression (RE) in comparison with vehicle-treated cells. and share the same β-Actin blots for p-mTor and RAS in both A549 and A549-TR cells.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: Western Blot, Expressing, Comparison

Journal: Cancers

Article Title: Oncogene Downregulation by Mahanine Suppresses Drug-Sensitive and Drug-Resistant Lung Cancer and Inhibits Orthotopic Tumor Progression

doi: 10.3390/cancers16213572

Figure Lengend Snippet: Effect of mahanine (MH) on the growth of orthotopic lung tumor growth. The line diagram depicts the experimental plan ( A ) and the effect of MH on tumorous lung weight ( B ), tumor progression in terms of luciferase signals ( C ), and growth inhibition ( D ) in female NOD Scid mice. Animals were inoculated with human Bioware Brite A549 Red-Luc cells (1.5 × 10 6 cells) in 50 μL Matrigel via intrathoracic injection to develop orthotopic tumors. Ten days post-implantation, animals were imaged, randomly assigned to treatment groups, and administered intraperitoneal injections of either the vehicle or MH (25 mg/kg) three times per week. Tumor growth kinetics were monitored via bioluminescent imaging twice weekly. Prior to each imaging session, animals received an intraperitoneal injection of luciferin substrate (80 mg/kg) 15 min before imaging. Tumor burden was quantified in vivo using a Biospace Lab Imager. At the study’s endpoint, animals were euthanized, and the weights of excised tumor-bearing lungs were recorded. Data are expressed as mean ± SD, and statistical analysis was conducted using Student’s t -test; ** p < 0.01; *** p < 0.001.

Article Snippet: Animals were inoculated with Bioware ® Brite A549 Red Fluc cells (1.5 × 10 6 cells) in 50 μL of Matrigel (BD Bioscience, Bedford, MA, USA) mixed with serum-free media (1:1; v / v ) via intrathoracic injection using 30-gauge needles, as described previously [ ].

Techniques: Luciferase, Inhibition, Injection, Imaging, In Vivo