Group |
Chemotherapy |
|
|||||
Before |
After |
D-value |
Before |
After |
D-value |
||
Symptoms |
|
|
|
|
|
|
|
Cough |
2.20±0.52 |
1.10±0.55** |
1.10±0.79 |
2.30±0.47 |
0.50±0.51** |
1.80±0.77△△ |
|
Phlegm blood |
2.35±0.49 |
1.60±0.60** |
0.75±0.72 |
2.45±0.51 |
0.70±0.47** |
1.75±0.44△△ |
|
Breath shortness |
2.20±0.62 |
1.00±0.73** |
1.20±0.89 |
2.30±0.73 |
0.55±0.60** |
1.75±0.97△ |
|
Chest/waist pain |
2.35±0.48 |
1.40±0.50** |
0.95±0.76 |
2.15±0.67 |
0.55±0.51** |
1.60±0.75△△ |
|
Expectoration |
2.55±0.51 |
1.45±0.60** |
1.10±0.55 |
2.30±0.66 |
0.50±0.51** |
1.80±0.70△△ |
|
Mental fatigue |
2.65±0.49 |
1.75±0.64** |
0.90±0.72 |
2.70±0.47 |
0.60±0.50** |
2.00±0.65△△ |
|
Mouth thirsty |
2.45±0.51 |
1.30±0.57** |
1.15±0.59 |
2.40±0.50 |
0.65±0.49** |
1.75±0.72△△ |
|
Night sweats |
2.55±0.51 |
1.35±0.49** |
1.20±0.52 |
2.81±0.41 |
0.70±0.57** |
2.10±0.64△△ |
|
Total scores |
19.30±2.49 |
10.95±1.28** |
8.35±2.49 |
19.30±2.66 |
4.75±1.48** |
14.55±2.50△△ |
|
Biomarkers |
|
|
|
|
|
|
|
ALT (U/L) |
27.25±8.96 |
33.30±7.78* |
-6.05±3.71 |
27.55±8.26 |
29.15±7.98 |
-1.60±2.04△△ |
|
Cr (μMol/L) |
59.95±22.52 |
67.85±19.53 |
-7.90±8.29 |
60.00±20.07 |
62.30±18.90 |
-3.70±7.38△ |
|
CEA (ng/mL) |
35.25±11.99 |
26.50±11.25* |
8.75±6.58 |
36.80±10.89 |
20.20±5.76** |
16.60±8.85△△ |
|
NSE (ng/mL) |
18.65±5.82 |
15.03±4.00* |
3.62±2.44 |
18.58±5.81 |
14.23±5.17* |
4.35±2.51 |
|
CYFRA21-1(ng/ml) |
4.08±1.39 |
3.32±1.32 |
0.76±0.67 |
4.03±1.35 |
3.13±1.15 |
0.90±1.04 |
|
TK1 (ng/mL) |
2.68±0.69 |
2.10±0.47* |
0.58±0.35 |
2.79±0.65 |
2.32±0.63* |
0.47±0.33 |
|
Compared with the before treatment group, *P < 0.05, **P < 0.01; Compared with the chemotherapy group, ? P < 0.05, ? ? P <0.01.br />
ALT, alanine transaminase; CEA, carcinoembryonic antigen; CR, creatinine; CYFRA21-1, cytokeratin fragment; NSE, neurone specific enolase; TK1, thymidine kinase 1; YKF, Yangfei Kongliu Formula
Table 1: The improvement of clinical symptom score and biomarker alteration before and after treatment x (± SD)
Figure 1: YKF promoting lung cancer A549 cell autophagy.
A, the CCK-8 viability after A549 cells treated with 5 ×, 10 ×, or 15 × YKF-contained serum for 48 h; B, A549 cells with 5 × YKF-contained serum for different times; C, the LC3 I, LC3 II, P62, and Beclin-1 protein by Western blotting; D, the relative ratio of LC3II/I; and E, the relative ratio of Beclin-1. YKF, Yangfei Kongliu Formula ***P < 0.001 compared with control.
Figure 2: YKF-induced autophagy or 3-MA inhibition on A549 cells.
A: the control electron photomicrographs of A549 cells (magnification, 13000 ×); B: the electron photomicrographs of autophagosomes in A549 cells treated with YKF for 48 h (magnification, 13000 ×); C: the pretreated A549 cells with or without 1 mM 3-MA exposed to YKF for an additional 48 h (magnification, 800 ×), and LC3 puncta were carried out by immunofluorescence technique. GFP, green fluorecent protein; 3-MA, 3-methyladenine; RFP, red fluorecent protein; YKF, Yangfei Kongliu Formula.
Figure 3: Silencing Beclin-1 reversed autophagy promoting effect of YKF on cancer cells.
A, the Beclin-1 iRNA plasmids were transfected to A549 cells and the si-Beclin-1-2 plasmid was screened with specific inhibiting Beclin-1 expression; B, the effects of si-Beclin-1-2 and 3-MA on cell viability of the A549 cells treated with YKF; C, the effects of si-Beclin-1-2 and 3-MA on cell autophagy related-protein expression of P62 and LC3B I/II. **P < 0.01, ***P < 0.001 compared with blank. ###P < 0.001 compared with 5 × YKF. &&&P < 0.001 compared with 5 × YKF + si-NC. 3-MA, 3-methyladenine; YKF, Yangfei Kongliu Formula
.JPG)
.JPG)
Figure 4: Silencing Beclin-1 reverses YKF inhibiting cell migration or invasion.
Representative photomicrographs of migratory cells (A ~ F) or invaded cells (A1 ~ F1) cells were counted in three independent experiments (n = 3). A, the A549 cells as a blank control; B, the A549 cells with YKF; C, the A549 cells with YKF plus specific si-Beclin-1-2; D, the A549 cells with YKF plus si-Beclin-1 negative control; E, the A549 cells with YKF plus 3-MA; F, the comparative analysis of A549 migratory cell numbers among the different groups; A1, the A549 cells as a blank control; B1, the A549 cells with YKF; C1, the A549 cells with YKF plus specific si-Beclin-1-2; D1, the A549 cells with YKF plus si-Beclin-1 negative control; E1, the A549 cells with YKF plus 3-MA; F1, the comparative analysis of A549 invaded cell numbers among the different groups. *P < 0.05, ***P < 0.001 compared with blank. ##P < 0.01, #P < 0.05 compared with 5 × YKF. 3-MA, 3-methyladenine; YKF, Yangfei Kongliu Formula.
Combined YKF
Tables at a glance
Figures at a glance