Application of LncRNA as biomarker and therapeutic target for non-small cell lung cancer

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to application of LncRNA as a non-small cell lung cancer biomarker and treatment target, wherein the nucleotide sequence of the LncRNA is shown as SEQ ID NO. 1. Experimental results of the application show that inhibition of expression of the LncRNA can prevent proliferation, survival and migration of non-small cell lung cancer cells, and therefore the LncRNA can be used as a non-small cell lung cancer biomarker and treatment target.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of a lncRNA as a biomarker and therapeutic target for non-small cell lung cancer. Background Technology

[0002] Lung cancer is the leading cause of cancer-related morbidity and mortality in my country and globally. If lung cancer is treated surgically in its early stages, the 5-year survival rate can exceed 50%. However, effective early diagnostic methods are currently lacking, and most lung cancer patients are diagnosed at an advanced stage with metastasis, at which point their sensitivity to radiotherapy and chemotherapy decreases. Although targeted therapy and immunotherapy targeting genes such as EGFR, ALK, ROS1, and MET have made some progress, the overall treatment outcome remains unsatisfactory, with a 5-year survival rate of only about 15% for advanced-stage patients. Therefore, identifying more biomarkers for early lung cancer screening and diagnosis, exploring potential therapeutic targets, and promoting the development of novel comprehensive treatment modalities are of significant clinical importance for improving the prognosis of lung cancer patients.

[0003] ncRNAs include short non-coding RNAs such as miRNAs, tsRNAs, and siRNAs, and long non-coding RNAs (lncRNAs). LncRNAs are a class of non-coding RNAs longer than 200 nt, lacking protein-coding ability or with limited coding ability. The GENCODE database shows that the human genome can generate more than 16,000 lncRNA genes, and some studies have shown that it can generate more than 100,000 lncRNAs. The abundance of lncRNAs provides a rich resource for targeted cancer therapy. LncRNAs play important regulatory roles in gene activation and silencing, X chromosome inactivation, alternative splicing, and post-translational regulation. Increasing research indicates that lncRNA expression is abnormal in various cancers, such as colorectal cancer, gastric cancer, breast cancer, and other types of tumors.

[0004] In the field of non-small cell lung cancer (NSCLC) research, there are no reports in the literature regarding the use of BCAN-AS2 as a diagnostic biomarker and therapeutic target. Therefore, developing BCAN-AS2 as a biomarker and therapeutic target for NSCLC will have significant clinical value and social implications for the early diagnosis and treatment of NSCLC. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides an application of lncRNA as a biomarker and therapeutic target for non-small cell lung cancer (NSCLC). Inhibiting the expression level of this lncRNA can prevent the proliferation, survival, and migration of NSCLC cells; therefore, this lncRNA can serve as a biomarker and therapeutic target for NSCLC.

[0006] The present invention solves the above-mentioned technical problems through the following technical solutions.

[0007] The present invention provides a LncRNA, wherein the LncRNA is LncRNA BCAN-AS2; the nucleotide sequence of the LncRNA BCAN-AS2 is shown in SEQ ID NO. 1.

[0008] This invention provides an application of the above-mentioned LncRNA as a biomarker in the preparation of products for diagnosing lung cancer.

[0009] In this invention, the lung cancer referred to is non-small cell lung cancer.

[0010] The present invention also provides the application of the above-mentioned LncRNA as a therapeutic target in the preparation of drugs for the prevention and / or treatment of lung cancer.

[0011] In this invention, the drug comprises an inhibitor that can inhibit the expression of the LncRNA.

[0012] In this invention, the drug comprises siRNA that targets the aforementioned LncRNA.

[0013] In this invention, the siRNA molecule is siRNA 1 or siRNA 2, wherein: the sense strand of siRNA 1 contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.2, and the antisense strand contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.3.

[0014] The sense strand of the siRNA 2 contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 4, and the antisense strand contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 5.

[0015] In this invention, the drug's effects include one or more of the following: proliferation, survival, and migration of lung cancer cells.

[0016] The present invention provides an siRNA molecule, wherein the siRNA molecule is siRNA 1 or siRNA 2, wherein: the sense strand of the siRNA 1 contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.2, and the antisense strand contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.3.

[0017] The sense strand of the siRNA 2 contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 4, and the antisense strand contains a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 5.

[0018] The present invention provides a composition comprising the above-described siRNA molecule.

[0019] This invention provides the use of the above-mentioned siRNA molecule or the above-mentioned pharmaceutical composition in the preparation of a drug for treating lung cancer.

[0020] In this invention, the lung cancer referred to is non-small cell lung cancer.

[0021] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0022] The reagents and raw materials used in this invention are all commercially available.

[0023] The positive and progressive effects of this invention are as follows: This invention has discovered that inhibiting the expression level of a certain LncRNA can prevent the proliferation, survival and migration of non-small cell lung cancer cells, and this LncRNA can serve as a biomarker and therapeutic target for non-small cell lung cancer. Attached Figure Description

[0024] Figure 1 Differentially expressed lncRNAs were screened from clinical samples using high-throughput sequencing; among them, Figure 1 (A) is a volcano diagram of LncRNA transcriptome sequencing; Figure 1 (B) is a transcriptome sequencing heatmap.

[0025] Figure 2 BCAN-AS2, a biomarker for lung cancer diagnosis, was analyzed using the TCGA dataset; among which, Figure 2 (A) represents the analysis results of unpaired samples. Figure 2 (B) shows the results of the paired sample analysis.

[0026] Figure 3 The results are from quantitative real-time PCR detection of BCAN-AS2 mRNA expression levels.

[0027] Figure 4 To download patient survival information from RNA-seq data of lung cancer tissue from the TCGA database, including... Figure 4 (A) is the overall survival curve (OS) for the patient's survival information. Figure 4 (B) is the recurrence-free survival curve (RFS) of the patient.

[0028] Figure 5 The effect of siRNA1 and siRNA2 knockdown on BCAN-AS2 is shown, where *** indicates p<0.001.

[0029] Figure 6 The effects of siRNA1 and siRNA2 on the inhibition of proliferation of lung cancer cells A549 and H1650.

[0030] Figure 7The effects of siRNA1 and siRNA2 on the survival of lung cancer cells A549.

[0031] Figure 8 The effects of siRNA1 and siRNA2 on the inhibition of migration in lung cancer cells A549 and H1650. Detailed Implementation

[0032] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0033] The BCAN-AS2 mentioned in this invention is located on chromosome 1 (Gene: ENSG00000229953.1, Position: Chromosome 1: 156,616,299-156,631,216), and its nucleotide sequence is shown in SEQ ID NO.1.

[0034] Example 1: High-throughput sequencing screening in clinical samples can serve as LncRNA biomarkers for lung cancer diagnosis.

[0035] In this embodiment, tumor tissues and adjacent normal tissues from five lung cancer patients were collected from Sir Run Run Shaw Hospital, affiliated with Zhejiang University School of Medicine. This study was approved by the Ethics Committee of Sir Run Run Shaw Hospital, affiliated with Zhejiang University School of Medicine. RNA was extracted and a transcriptome sequencing library was constructed. Differentially expressed lncRNAs were screened using high-throughput sequencing. The transcriptome sequencing volcano plot of the lncRNAs was obtained, as shown in the figure. Figure 1 (A) shows a transcriptome sequencing heatmap, as shown below. Figure 1 As shown in (B).

[0036] according to Figure 1 (A) It can be seen that 283 LncRNA molecules were upregulated in tumors, while 180 LncRNAs were downregulated. Among them, the average expression value of BCAN-AS2 in tumors was 14.2, which was 17.1 times upregulated compared to the average expression value of 0.83 in normal tissues.

[0037] Experimental results demonstrate that BCAN-AS2 expression is significantly upregulated in lung cancer tissues, indicating its potential as a diagnostic biomarker for lung cancer.

[0038] Example 2: Big data analysis of BCAN-AS2 as a biomarker for lung cancer diagnosis.

[0039] This embodiment, based on the TCGA database, systematically analyzed the mRNA expression characteristics of BCAN-AS2 in lung cancer tissues by comparing transcriptome sequencing data from 59 adjacent normal tissues and 526 lung cancer tumor tissues. The analysis results are as follows: Figure 2 As shown, the analysis results of unpaired samples are as follows: Figure 2 As shown in (A), the results of the paired sample analysis are as follows: Figure 2 As shown in (B).

[0040] according to Figure 2 (A) It can be seen that the average expression value of BCAN-AS2 in the 59 unpaired adjacent normal tissue samples was 2.08, while the average expression value in the 526 unpaired lung cancer tumor tissue samples was 4.76, with a p value of 2.1e-38, which is statistically significant.

[0041] according to Figure 2 (B) It can be seen that in the 58 paired adjacent normal tissue samples and tumor tissue samples, the mean expression values ​​of BCAN-AS2 were 2.08 and 4.53, respectively, with a p value of 2.55e-13, which is statistically significant.

[0042] The above analysis results indicate that BCAN-AS2 expression in lung cancer tumor tissues is significantly higher than that in adjacent normal tissues, suggesting that BCAN-AS2 expression is significantly upregulated in lung cancer patients. The results of this study are consistent with the results of the clinical sequencing data in Example 1.

[0043] Example 3: Validating BCAN-AS2 as a biomarker for lung cancer diagnosis.

[0044] This embodiment aims to further verify the expression of BCAN-AS2 in lung cancer samples. The specific steps are as follows: Sample collection: Surgical tumor tissue and adjacent normal tissue were collected from 55 patients diagnosed with lung cancer at Sir Run Run Shaw Hospital, affiliated with Zhejiang University School of Medicine. Each patient was informed of the purpose of the sample and signed an informed consent form. The samples were removed and stored in liquid nitrogen. This study was approved by the Ethics Committee of Sir Run Run Shaw Hospital, affiliated with Zhejiang University School of Medicine.

[0045] RNA extraction from tissue: A suitable amount of tissue was placed in a 1.5 mL centrifuge tube, 1 mL of Trizol reagent and a steel ball were added, and homogenized using a homogenizer. After homogenization, 200 μL of chloroform was added, and the mixture was vortexed for 15 seconds to mix. The mixture was then allowed to stand at room temperature for 5 minutes. Centrifuged at 12000 g for 5 minutes at 4°C, and 500 μL of the supernatant was carefully transferred to a new centrifuge tube. An equal volume of isopropanol was added, and the mixture was allowed to stand on ice for 10 minutes. Centrifuged at 12000 g for 10 minutes at 4°C, and the supernatant was discarded. The tissue was washed with 75% ethanol, centrifuged at 8000 g at 4°C, and the supernatant was discarded. Step 5 was repeated once. The centrifuge tubes were then air-dried in a fume hood for 10 minutes. The RNA samples were then dissolved in an appropriate amount of DEPC water. The Nanodrop concentration was determined, and the RNA samples were stored frozen at -80°C.

[0046] Reverse transcription and quantitative detection: Reverse transcription was performed using the HiScript® II Q RT SuperMix for qPCR (+gDNA wiper) reverse transcription kit from Nanjing Novizan Biotechnology Co., Ltd., according to the instructions. Then, the RT-qPCR quantitative system was prepared using the company's ChamQ Universal SYBR qPCR Master Mix (Q711) kit. Finally, the expression in the samples was detected on a Bio-Rad CFX-96 real-time PCR instrument.

[0047] The results of the real-time PCR test are as follows: Figure 3 As shown in the figure. Specifically, the expression of BCAN-AS2 in tumor samples was significantly higher than that in paired adjacent normal tissue samples, with a p-value of less than 0.0001, which was statistically significant.

[0048] Experimental results demonstrated that BCAN-AS2 expression in lung cancer tumor tissues was significantly higher than in adjacent normal tissues. Therefore, the experimental results further successfully validated that BCAN-AS2 can serve as a diagnostic biomarker for lung cancer.

[0049] Example 4: Assessing the relationship between BCAN-AS2 expression levels and prognosis in lung cancer patients.

[0050] Assessing BCAN-AS2 expression and prognosis in lung cancer patients: RNA-seq data of lung cancer tissues were downloaded from the TCGA database, and overall survival (OS) and recurrence-free survival (RFS) curves were plotted in conjunction with patient survival information. The analysis results are as follows: Figure 4 As shown.

[0051] By analyzing the overall survival time of 252 patients with high BCAN-AS2 expression and 250 patients with low expression, the overall survival curves (OS) of the patients' survival information are as follows: Figure 4 As shown in (A), the results indicate that the overall survival time of patients with high expression was significantly shorter than that of patients with low expression.

[0052] By analyzing the recurrence-free survival time of 135 lung cancer patients with high BCAN-AS2 expression and 163 lung cancer patients with low BCAN-AS2 expression, the recurrence-free survival curves (RFS) of the patients were obtained as follows: Figure 4 As shown in (B), the results indicate that patients with high expression had a shorter relapse-free survival than those with low expression.

[0053] The above analysis results indicate that lung cancer patients with high BCAN-AS2 expression have significantly shorter overall survival and recurrence-free survival than those with low expression. This suggests that high BCAN-AS2 expression predicts a worse prognosis for lung cancer patients, indicating a significant negative correlation between BCAN-AS2 expression levels and patient survival time.

[0054] Therefore, BCAN-AS2 can serve as an effective biomarker for lung cancer prognosis assessment and has the function of clinical diagnosis of lung cancer.

[0055] Example 5: The effect of interfering with BCAN-AS2 expression on lung cancer.

[0056] To investigate the biological function of BCAN-AS2 in the development and progression of lung cancer, we used RNA interference technology to specifically knock down BCAN-AS2 expression in human lung cancer cell lines. First, we verified the interference efficiency using real-time quantitative PCR. Then, we evaluated the function of BCAN-AS2 in influencing lung cancer cell proliferation and survival using the CCK-8 assay and colony formation assay system. Finally, we monitored its therapeutic effect on lung cancer by detecting the effect of BCAN-AS2 on lung cancer cell migration using the trans-well assay. Specifically, the lung cancer cells A549 and H1650 used in this example were preserved in the laboratory. We designed siRNA1, siRNA2, and siCrtl to specifically target and knock down BCAN-AS2. The sense and antisense sequences of siRNA1, siRNA2, and siCrtl are shown in Table 1 below. The siRNA1, siRNA2, and siCrtl used in this example were synthesized by Shanghai Jima Biotechnology Co., Ltd.

[0057] Table 1:

[0058] The general experimental steps are as follows: Cell resuscitation and culture: 1. Remove the cells from liquid nitrogen and quickly place them in a 37°C constant temperature water bath to thaw. After fully thawing, centrifuge the cells at 800 rpm and discard the old culture medium.

[0059] 2. Then add 1 mL of 1640 complete medium containing 10% FBS and antibiotics to resuspend the cells. Add the resuspended cells to a 10 cm cell culture dish and add an additional 7 mL of 1640 medium.

[0060] 3. Place the revived cells in a cell culture incubator (37°C, containing 5% CO2), and replace with fresh culture medium the next day. Change the culture medium every two days, and passage the cells every three days.

[0061] siRNA transfection: 1. Dissolve the siRNA powder synthesized by Gemma Biotechnology in RNase-free water to a concentration of 10 μM, aliquot into 10 μL and store at -20℃.

[0062] 2. Then, A549 and H1650 cells in the logarithmic growth phase were seeded into 6-well plates. After 18-24 hours of seeding, the cell confluence reached 60%-70%. GeneMute was then used to... TM Transfection was performed using the transfection reagent. 30 minutes before transfection, each well was replaced with 1 mL of fresh medium containing 10% FBS. The transfection system is as follows:

[0063]

[0064] Mix well by blowing and incubate at room temperature for 15 minutes.

[0065] 3. Add 100 μL of the mixed transfection reagent to a 6-well plate, mix well, and incubate at 37°C in a 5% CO2 incubator for 24 hours before proceeding with subsequent experiments.

[0066] RNA extraction: 1. After siRNA transfection for 48 hours, discard the culture medium, wash twice with PBS, and add 1 mL of TRIZOL reagent to lyse the cells.

[0067] 2. Then transfer the lysis buffer to a 1.5 mL centrifuge tube, add 200 μL of chloroform, vortex for 15 seconds to mix, and let stand at room temperature for 5 minutes.

[0068] Centrifuge at 12000g for 5 minutes at 4℃. Carefully aspirate 500μL of the supernatant into a new centrifuge tube, add an equal volume of isopropanol, and incubate on ice for 10 minutes.

[0069] 4. Centrifuge at 12000g for 10 minutes at 4℃, and discard the supernatant.

[0070] 5. Wash with 75% ethanol, centrifuge at 8000g at 4℃, and discard the supernatant.

[0071] 6. Repeat step 5 once, then open the centrifuge tubes and air dry in a fume hood for 10 minutes. Then add an appropriate amount of DEPC in water to dissolve.

[0072] 7. Nanodrop concentration determination, frozen storage at -80℃.

[0073] RT-qPCR detection of BCAN-AS2 knockdown efficiency: Reverse transcription was performed using the HiScript® II Q RTSuperMix for qPCR (+gDNA wiper) (R233) kit from Nanjing Novizan Pharmaceutical Co., Ltd.

[0074] 1. Remove genomic DNA and prepare a mixture in a 200 μL centrifuge tube according to the table below:

[0075] 2. Gently pipette to mix, and incubate at 42°C for 2 minutes to degrade DNA.

[0076] 3. Configure the reverse transcription reaction system by directly adding 5×HiScript II qRTSuperMix II to the reaction tube from step 1:

[0077] 4. After mixing with a pipette, perform reverse transcription according to the following steps: 50℃ for 15 minutes, 85℃ for 5 seconds, and 4℃ for 1 hour. The product can be used immediately for qPCR after the reaction is complete, or stored at -20℃.

[0078] 5. RT-qPCR quantitative reaction was performed using ChamQ Universal SYBR qPCR Master Mix (Q711) reagent from Nanjing Novizan Pharmaceutical Co., Ltd. The reaction system is as follows:

[0079] Reaction program settings: 1 cycle of pre-denaturation (95℃, 30 seconds), followed by 40 cycles of {denaturation (95℃, 10 seconds), annealing / extension (60℃, 30 seconds)}, using the default melting curve on the qPCR instrument. After quantification, export the experimental data.

[0080] Use 2 -ΔΔCt The expression level of BCAN-AS2 was calculated using the method described below, and the results are shown in Table 2.

[0081] Table 2:

[0082] In this embodiment, BCAN-AS2 was knocked down using siRNA, and the knockdown effect was verified by RT-qPCR. The experimental results are as follows: Figure 5 As shown.

[0083] according to Figure 5 It is evident that by designing specific siRNAs targeting BCAN-AS2 and then knocking down BCAN-AS2 in tumor cells, quantitative real-time PCR results show that both siRNAs can effectively interfere with BCAN-AS2 expression. This indicates that the siRNAs of this invention can be used as drugs for treating lung cancer.

[0084] CCK-8 assay to detect the effect of interfering with BCAN-AS2 expression on lung cancer cell proliferation.

[0085] 1. 24 hours after cell transfection, the cells were digested with 0.25% trypsin and counted using a hemocytometer.

[0086] 2. Seed 1000-1500 cells per well in a 96-well plate, with 5 replicates and 6 time points (0, 24, 48, 72, 96, 120 hours) per group.

[0087] 3. After cell adhesion, add 10 μL of CCK-8 reagent to the 0-hour cells, incubate at 37°C in the dark for 2 hours, and then measure the absorbance (OD) value at a wavelength of 450 nm.

[0088] 4. Then, add CCK-8 reagent every 24 hours and measure the absorbance OD value.

[0089] The effect of siRNA interference with BCAN-AS2 expression on the proliferation of lung cancer A549 cells is shown in Table 3 below.

[0090] Table 3:

[0091] The effect of siRNA interference with BCAN-AS2 expression on the proliferation of lung cancer H1650 cells is shown in Table 4 below.

[0092] Table 4:

[0093] according to Figure 6 It was found that knocking down BCAN-AS2 significantly inhibited the proliferation rate of lung cancer cells A549 and H1650. This suggests that BCAN-AS2 may be involved in the development and progression of lung cancer and could serve as a novel therapeutic target.

[0094] To investigate the effect of interfering with BCAN-AS2 expression on lung cancer cell survival.

[0095] 1. Cells transfected with BCAN-AS2 siRNA were digested with trypsin and counted using a hemocytometer.

[0096] 2. Seed cells in 12-well plates at a density of 800 cells / well.

[0097] 3. Incubate the cell culture plates at 37°C in a 5% CO2 incubator for 7-8 days. Observe under a microscope. When cells form distinct and independent clonal plaques, discard the culture medium, wash once with PBS, air dry, add 500 μL of methanol to each well, and fix for 10 minutes.

[0098] 4. Discard the methanol, let it dry, add 500 μL of 0.1% crystal violet solution to each well, and stain for 10 minutes.

[0099] Discard the crystal violet solution and wash each well twice with 500 μL of PBS. Take photos of the results, as shown below. Figure 7 As shown.

[0100] from Figure 7 The experimental results show that knocking down BCAN-AS2 in lung cancer A549 cells using siRNA and detecting cell survival through a colony formation assay demonstrates that knocking down BCAN-AS2 expression significantly inhibits lung cancer cell survival, with siRNA showing the best effect in inhibiting lung cancer cell survival.

[0101] To investigate the effect of interfering with BCAN-AS2 expression on lung cancer cell migration.

[0102] 1. After cell plating and transfection with BCAN-AS2-specific siRNA for 24 hours, lung cancer cells were starved in serum-free medium for 24 hours.

[0103] 2. After starvation, discard the culture medium, wash once with PBS, digest the cells with 0.25% trypsin, then stop the digestion with complete culture medium, centrifuge to remove the supernatant, and then resuspend the cells in FBS-free culture medium and count the cells using a hemocytometer.

[0104] 3. Add 500 μL of culture medium containing 10% FBS to a 24-well plate, and place the Transwell chamber into the plate. Follow the 1×10⁻⁶ cycle. 5 Prepare a cell suspension at a density of 100 cells per milliliter, and slowly add 300 μL to the upper layer of the Transwell chamber. Incubate at 37°C in a 5% CO2 incubator for 18-24 hours.

[0105] 4. Remove the chamber, discard the remaining culture medium, air dry, and then fix with methanol for 10 minutes.

[0106] 5. After fixation, discard excess methanol, air dry the chamber, and then stain the chamber in 0.1% crystal violet solution for 10 minutes.

[0107] 6. Discard the crystal violet, wash twice with PBS, and wipe away any unmigrated cells in the upper chamber with a cotton swab. Under an upright microscope, randomly select three fields of view to photograph and count the cells. Take the average value to represent the cell migration ability. The photographic results are shown below. Figure 8 As shown.

[0108] according to Figure 8 The experimental results showed that knocking down BCAN-AS2 in lung cancer A549 and H1650 cells and detecting cell migration ability by trans-well assays indicated that interfering with BCAN-AS2 expression could significantly inhibit tumor cell migration.

Claims

1. The application of a lncRNA as a therapeutic target in the preparation of drugs for the prevention and / or treatment of non-small cell lung cancer, wherein, The LncRNA is LncRNA BCAN-AS2; the nucleotide sequence of LncRNA BCAN-AS2 is shown in SEQ ID NO. 1; The drug contains siRNA that targets the aforementioned LncRNA, wherein the siRNA molecule is siRNA 2, wherein: The sense strand of the siRNA 2 is a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 4, and the antisense strand is a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.

5.

2. A siRNA molecule, wherein the siRNA molecule is siRNA 2, wherein: The sense strand of the siRNA 2 is a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 4, and the antisense strand is a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO.

5.

3. The use of the siRNA molecule as described in claim 2 in the preparation of a drug for treating non-small cell lung cancer.

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