CN112813163A - Marker for myocardial injury in gastric cancer chemotherapy - Google Patents
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- 208000005718 Stomach Neoplasms Diseases 0.000 title claims abstract description 56
- 238000002512 chemotherapy Methods 0.000 title claims abstract description 56
- 206010017758 gastric cancer Diseases 0.000 title claims abstract description 56
- 201000011549 stomach cancer Diseases 0.000 title claims abstract description 56
- 239000003550 marker Substances 0.000 title claims abstract description 13
- 208000037891 myocardial injury Diseases 0.000 title claims description 11
- 230000003680 myocardial damage Effects 0.000 claims abstract description 40
- 238000003745 diagnosis Methods 0.000 claims abstract description 12
- 238000001514 detection method Methods 0.000 claims abstract description 11
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- 238000004393 prognosis Methods 0.000 claims description 8
- 210000004369 blood Anatomy 0.000 claims description 6
- 239000008280 blood Substances 0.000 claims description 6
- 238000009007 Diagnostic Kit Methods 0.000 claims description 5
- 210000005259 peripheral blood Anatomy 0.000 abstract description 22
- 239000011886 peripheral blood Substances 0.000 abstract description 22
- 238000013399 early diagnosis Methods 0.000 abstract description 3
- 208000024172 Cardiovascular disease Diseases 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000003753 real-time PCR Methods 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 208000029549 Muscle injury Diseases 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
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- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 208000013875 Heart injury Diseases 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
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- 238000000034 method Methods 0.000 description 2
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Abstract
The invention provides a marker for myocardial damage caused by gastric cancer chemotherapy, belonging to the technical field of cardiovascular diseases. The invention proves that LncRNA-RP11-114O8.1 is remarkably low expressed in peripheral blood of a patient with gastric cancer myocardial damage, so that the LncRNA-RP11-114O8.1 can be used for auxiliary diagnosis of the patient with gastric cancer myocardial damage. Compared with a biochemical detection method, the fluorescence quantitative PCR detection method has higher sensitivity and specificity, and is beneficial to realizing early diagnosis and early treatment of the patients with the myocardial damage caused by the gastric cancer chemotherapy.
Description
Technical Field
The invention belongs to the technical field of cardiovascular diseases, and particularly relates to a marker for myocardial damage related to gastric cancer chemotherapy.
Background
Gastric cancer is a highly lethal disease, the most common type of tumor in the digestive tract. Currently, over 70 million people die of gastric cancer every year, and nearly 100 million new-onset gastric cancer patients every year. At present, the main treatment modes of the gastric cancer are surgical treatment, radiotherapy, chemotherapy, targeted treatment and the like. Among them, chemotherapy plays a key role in the treatment of patients with advanced gastric cancer, and the treatment effect is significant. However, chemotherapy treatment can lead to irreversible cardiotoxicity in patients, and ultimately patients can affect their survival due to heart failure and severe arrhythmia. Therefore, the early diagnosis of the myocardial damage related to the chemotherapy of the gastric cancer patient can effectively improve the survival rate of the patient.
LncRNA refers to endogenous non-protein-encoding transcripts greater than 200 nucleotides in length. With the development of high throughput sequencing technology, researchers have found that deregulated expression of lncRNA alters disease progression and can serve as an independent biomarker for a variety of diseases. The lncRNA can be directly detected from the plasma and serum of a patient, so the method has the advantages of convenience and rapidness in detection, high sensitivity, high specificity and no wound. Therefore, the study of the myocardial injury lncRNA associated with gastric cancer chemotherapy helps to realize early diagnosis and early treatment of the myocardial injury of patients.
Disclosure of Invention
The invention aims to provide a marker for myocardial damage caused by gastric cancer chemotherapy.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a marker LncRNA-RP11-114O8.1 for auxiliary diagnosis and prognosis diagnosis of gastric cancer chemotherapy myocardial injury, wherein the sequence of LncRNA-RP11-114O8.1 is shown as SEQ ID No. 1.
In addition, the invention provides application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing an auxiliary diagnostic kit for myocardial damage caused by gastric cancer chemotherapy.
Preferably, the reagent is a detection reagent of LncRNA-RP11-114O8.1 specific primers.
Preferably, the sequence of the upstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO.2, and the sequence of the downstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO. 3.
In addition, the invention provides application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing a kit for prognosis diagnosis of myocardial damage caused by gastric cancer chemotherapy.
Preferably, the reagent is a detection reagent of LncRNA-RP11-114O8.1 specific primers.
Preferably, the sequence of the upstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO.2, and the sequence of the downstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO. 3.
In addition, the invention provides application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing a diagnostic kit for distinguishing patients with myocardial damage caused by gastric cancer chemotherapy and patients without myocardial damage caused by gastric cancer chemotherapy.
Preferably, the reagent is a detection reagent of LncRNA-RP11-114O8.1 specific primers.
Preferably, the sequence of the upstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO.2, and the sequence of the downstream primer of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO. 3.
The invention has the beneficial effects that:
the invention discovers that the expression of LncRNA-RP11-114O8.1 in the peripheral blood of a patient with cardiac muscle injury caused by gastric cancer chemotherapy is obviously reduced compared with the expression in the peripheral blood of a patient with gastric cancer who is not treated by chemotherapy; meanwhile, the decrease is obviously different from the expression of LncRNA-RP11-114O8.1 in peripheral blood of a patient without myocardial damage in gastric cancer chemotherapy; in addition, after treatment, the expression quantity of LncRNA-RP11-114O8.1 in peripheral blood of a patient with cardiac injury caused by gastric cancer chemotherapy, the symptoms of which are relieved, is obviously improved compared with the peripheral blood of the patient with cardiac injury caused by gastric cancer chemotherapy, and the difference has statistical significance. Therefore, the LncRNA-RP11-114O8.1 can be used as a marker for auxiliary diagnosis and prognosis diagnosis of patients with myocardial damage caused by gastric cancer chemotherapy, and can be used as a marker for distinguishing patients with myocardial damage caused by gastric cancer chemotherapy and patients without myocardial damage caused by gastric cancer chemotherapy.
Drawings
FIG. 1 expression differences of LncRNA-RP11-114O8.1 in groups a, b, c and d,
a group: peripheral blood of a gastric cancer patient not treated by chemotherapy,
b group: peripheral blood of patients with gastric cancer chemotherapy myocardial damage,
and c, group: peripheral blood of patients without myocardial damage in gastric cancer chemotherapy,
and d, group: peripheral blood of a patient with gastric cancer chemotherapy myocardial injury, who has relieved symptoms of myocardial injury after treatment;
FIG. 2 ROC curves of difference in expression of LncRNA-RP11-114O8.1 in group a and group b;
FIG. 3 ROC curves of difference in expression of LncRNA-RP11-114O8.1 in group b and group c;
FIG. 4 ROC curves for differences in expression of LncRNA-RP11-114O8.1 in group b and group d.
Detailed Description
In order to clearly illustrate the technical features of the present solution, the present solution is explained below by way of specific embodiments.
Example 1
Detection sample selection
1. Grouping of test specimens
a group: 45 patients with gastric cancer had peripheral blood (not treated with chemotherapy),
b group: the peripheral blood of 35 patients with gastric cancer chemotherapy myocardial damage,
and c, group: 15 cases of patients with gastric cancer chemotherapy without myocardial damage peripheral blood,
and d, group: 16 cases of peripheral blood of patients with gastric cancer chemotherapy myocardial damage, with post-treatment myocardial damage remission;
2. extraction of peripheral blood serum RNA according to groups
(1) Placing peripheral blood of a patient in a centrifuge, centrifuging at 3000rpm/min for 15min, and transferring the serum of the supernatant into a new centrifuge tube;
(2) taking 250 ul of serum out, placing the serum in an EP tube, adding 750 ul of Trizol reagent, repeatedly beating and uniformly mixing, and standing for 5 min;
(3) adding 200 μ l chloroform, shaking the mixture for 15s, standing at room temperature for 5min, and centrifuging at 4 deg.C and 12000rpm for 15 min;
(4) transferring the supernatant to a new EP tube, adding isopropanol with the same amount as the supernatant, reversing and mixing uniformly, standing at room temperature for 10min, centrifuging at 4 ℃ at 12000rpm for 15min, and discarding the supernatant;
(5) adding 1ml of precooled 75% ethanol to resuspend the RNA precipitate, gently washing the precipitate, centrifuging at 4 ℃ and 7500rpm for 15min, removing supernatant, and standing at room temperature to remove ethanol on the tube wall;
(6) the concentration and purity of the extracted RNA was determined using a spectrophotometer.
3. Reverse transcription of RNA into cDNA
(1) Using PrimeScriptTMThe RT Master Mix kit is prepared by adding the following substances into a centrifugal tube without RNA enzyme to prepare a reverse transcription reaction system:
| composition (I) | Volume of |
| 5×PrimeScript Buffer | 4μl |
| PrimeScript RT Enzyme Mix I | 1μl |
| Oligo dT Primer | 1μl |
| Random 6 Primers | 1μl |
| Total RNA | 1μg |
| RNase Free dH2O | Make up to 20. mu.l |
(2) The reaction conditions were as follows:
15min at 37 ℃; 5s at 85 ℃; keeping at 4 ℃.
Real-time PCR detection of expression level of LncRNA-RP11-114O8.1
(1) The reaction was added as follows:
| composition (I) | Volume of |
| SYBR⑩Premix Ex Taq II | 10μl |
| PCR Forward Primer | 0.8μl |
| PCR Reverse Primer | 0.8μl |
| ROX Reference Dye | 0.4μl |
| DNA template | 2μl |
| dH2O | 6μl |
| Total | 20μl |
Wherein, the primer sequence of LncRNA-RP11-114O8.1 is as follows:
Forward Primer: 5’-CACTGCAACTTGCTTTGACCA-3’，SEQ ID NO.2；
Reverse Primer: 5’-CTGAGACATCTGGGTGAGGC-3’, SEQ ID NO.3；
(2) the reaction process is as follows:
pre-denaturation at 95 ℃ for 30 s; repeating 40 cycles at 95 ℃ for 5s and 60 ℃ for 34 s;
beta-actin is used as internal reference, 2 is adopted-∆∆CtThe method of (3) calculates the relative expression level of LncRNA-RP11-114O 8.1.
The experimental results are shown in FIG. 1, the relative expression amount of LncRNA-RP11-114O8.1 in group b is 0.479 + -0.038, the relative expression amount of LncRNA-RP11-114O8.1 in group c is 0.932 + -0.043, and the relative expression amount of LncRNA-RP11-114O8.1 in group d is 0.844 + -0.040;
the above results indicate that the relative expression level of LncRNA-RP11-114O8.1 in peripheral blood of patients with myocardial damage due to gastric cancer chemotherapy is significantly reduced (P < 0.0001) compared with that of patients without chemotherapy, while the relative expression level of LncRNA-RP11-114O8.1 in peripheral blood of patients with gastric cancer without myocardial damage after chemotherapy is not significantly different, which indicates that LncRNA-RP11-114O8.1 is low expressed in patients with myocardial damage due to gastric cancer chemotherapy, and the patients with myocardial damage can be detected by detecting the expression level of LncRNA-RP11-114O8.1 after chemotherapy.
Secondly, the relative expression level of LncRNA-RP11-114O8.1 in peripheral blood of a patient with cardiac muscle damage caused by gastric cancer chemotherapy and a patient without cardiac muscle damage caused by gastric cancer chemotherapy is obviously different (P < 0.0001), which indicates that whether the patient has the cardiac muscle damage can be effectively distinguished by detecting the relative expression level of LncRNA-RP11-114O8.1 in the peripheral blood of the patient after the chemotherapy.
Secondly, the relative expression level of LncRNA-RP11-114O8.1 in peripheral blood of a patient with remission of myocardial damage after treatment is remarkably improved (P < 0.0001) compared with that of a patient with myocardial damage caused by gastric cancer chemotherapy, which indicates that the relative expression level of LncRNA-RP11-114O8.1 in peripheral blood of a patient with remission of myocardial damage after treatment is remarkably different from that of a patient with myocardial damage after treatment, so that the LncRNA-RP11-114O8.1 can be used for prognosis diagnosis of a patient with myocardial damage caused by gastric cancer chemotherapy.
Example 2
For further investigation, whether LncRNA-RP11-114O8.1 can be used as a diagnostic and prognostic marker for chemotherapy-related myocardial injury, ROC curves between groups a and b, b and c, and b and d were calculated and plotted in the present invention, as shown in FIG. 2, FIG. 3, and FIG. 4.
As can be seen from FIG. 2, the AUC between group a and group b is 0.9152, Std. error is 0.03068, and P is < 0.0001, which indicates that LncRNA-RP11-114O8.1 has excellent diagnostic value in the auxiliary diagnosis of patients with myocardial damage caused by gastric cancer chemotherapy.
As can be seen from FIG. 3, the AUC values between group b and group c are 0.9419, Std. Error is 0.3073, and P is < 0.0001, indicating that LncRNA-RP11-114O8.1 has excellent diagnostic value in the differential diagnosis of patients with myocardial damage due to gastric cancer chemotherapy and patients without myocardial damage due to gastric cancer chemotherapy.
As can be seen from FIG. 4, the AUC between group b and group d is 0.8973, Std. error is 0.4357, and P is < 0.0001, which indicates that LncRNA-RP11-114O8.1 has excellent diagnostic value in the prognosis of patients with myocardial damage caused by gastric cancer chemotherapy.
Therefore, the LncRNA-RP11-114O8.1 can be used as a marker for auxiliary diagnosis and prognosis diagnosis of patients with myocardial damage caused by gastric cancer chemotherapy, and can be used as a marker for distinguishing patients with myocardial damage caused by gastric cancer chemotherapy and patients without myocardial damage caused by gastric cancer chemotherapy.
Sequence listing
<110> ninth people hospital in Qingdao city
<120> marker for myocardial damage caused by gastric cancer chemotherapy
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 191
<212> DNA
<213> Human source (Human)
<400> 1
atcacacacc gggacctgtc atgggttgac actgcaactt gctttgacca atagcatgga 60
cagaaatgag acctaccaac tttatattgc tctcttcgag gtctgtgcca ccacgtaaga 120
agtataacta tgctgcttct agtgaagttg agcctcaccc agatgtctca gctaccataa 180
ctaaggcact a 191
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<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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cactgcaact tgctttgacc a 21
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<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ctgagacatc tgggtgaggc 20
Claims (10)
1. A marker LncRNA-RP11-114O8.1 for auxiliary diagnosis and prognosis diagnosis of gastric cancer chemotherapy myocardial injury, wherein the sequence of LncRNA-RP11-114O8.1 is shown as SEQ ID No. 1.
2. Application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing an auxiliary diagnostic kit for myocardial injury in gastric cancer chemotherapy.
3. The use according to claim 2, wherein the reagent is a detection reagent for LncRNA-RP11-114O8.1 specific primers.
4. The use of claim 3, wherein the forward primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO.2, and the reverse primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO. 3.
5. Application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing a gastric cancer chemotherapy myocardial injury prognosis diagnostic kit.
6. The use of claim 5, wherein the reagent is a detection reagent for LncRNA-RP11-114O8.1 specific primers.
7. The use of claim 6, wherein the forward primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO.2, and the reverse primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID NO. 3.
8. Application of a reagent for detecting the expression level of LncRNA-RP11-114O8.1 in a blood sample in preparing a diagnostic kit for distinguishing patients with myocardial damage caused by gastric cancer chemotherapy and patients without myocardial damage caused by gastric cancer chemotherapy.
9. The use of claim 8, wherein the reagent is a detection reagent for LncRNA-RP11-114O8.1 specific primers.
10. The use of claim 9, wherein the forward primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID No.2, and the reverse primer sequence of the LncRNA-RP11-114O8.1 specific primer is shown as SEQ ID No. 3.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107419004A (en) * | 2017-04-28 | 2017-12-01 | 青岛大学 | LncRNA RP11 290F20.3 and its siRNA application |
| CN109609648A (en) * | 2019-01-31 | 2019-04-12 | 泰山医学院 | LncRNA marker relevant to liver cancer and its detection primer and application |
| CN110172514A (en) * | 2019-06-04 | 2019-08-27 | 中国人民解放军联勤保障部队第九六0医院 | For developing the molecular marker of sdenocarcinoma of stomach diagnosis product |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107419004A (en) * | 2017-04-28 | 2017-12-01 | 青岛大学 | LncRNA RP11 290F20.3 and its siRNA application |
| CN109609648A (en) * | 2019-01-31 | 2019-04-12 | 泰山医学院 | LncRNA marker relevant to liver cancer and its detection primer and application |
| CN110172514A (en) * | 2019-06-04 | 2019-08-27 | 中国人民解放军联勤保障部队第九六0医院 | For developing the molecular marker of sdenocarcinoma of stomach diagnosis product |
Non-Patent Citations (1)
| Title |
|---|
| BIRREN,B等: "Homo sapiens chromosome 8, clone RP11-114O8, complete sequence", 《NCBI》 * |
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